81 results on '"Nina N Nupponen"'
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2. The Peptide–Drug Conjugate Melflufen Modulates the Unfolded Protein Response of Multiple Myeloma and Amyloidogenic Plasma Cells and Induces Cell Death
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Ken Flanagan, Romika Kumari, Juho J. Miettinen, Staci L. Haney, Michelle L. Varney, Jacob T. Williams, Muntasir M. Majumder, Minna Suvela, Ana Slipicevic, Fredrik Lehmann, Nina N. Nupponen, Sarah A. Holstein, and Caroline A. Heckman
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Diseases of the blood and blood-forming organs ,RC633-647.5 - Abstract
Immunoglobulin light-chain (AL) amyloidosis is a rare disease caused by clonal plasma cell secretion of misfolded light chains that assemble as toxic amyloid fibrils, depositing in vital organs including the heart and kidneys, causing organ dysfunction. Plasma cell–directed therapeutics are expected to reduce production of toxic light chain by eliminating amyloidogenic cells in bone marrow, thereby diminishing amyloid fibril deposition and providing the potential for organ recovery. Melphalan flufenamide (melflufen) is a first-in-class peptide–drug conjugate that targets aminopeptidases and rapidly releases alkylating agents inside tumor cells. Melflufen is highly lipophilic, permitting rapid uptake by cells, where it is enzymatically hydrolyzed by aminopeptidases, resulting in intracellular accumulation of the alkylating agents, including melphalan. Previous data demonstrating sensitivity of myeloma cells to melflufen suggest that the drug might be useful in AL amyloidosis. We describe the effects of melflufen on amyloidogenic plasma cells in vitro and ex vivo, demonstrating enhanced cytotoxic effects in comparison to melphalan, as well as novel mechanisms of action through the unfolded protein response (UPR) pathway. These findings provide evidence that melflufen-mediated cytotoxicity extends to amyloidogenic plasma cells, and support the rationale for the evaluation of melflufen in patients with AL amyloidosis.
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- 2022
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3. Growth Response and Differentiation of Bone Marrow-Derived Mesenchymal Stem/Stromal Cells in the Presence of Novel Multiple Myeloma Drug Melflufen
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Arjen Gebraad, Roope Ohlsbom, Juho J. Miettinen, Promise Emeh, Toni-Karri Pakarinen, Mikko Manninen, Antti Eskelinen, Kirsi Kuismanen, Ana Slipicevic, Fredrik Lehmann, Nina N. Nupponen, Caroline A. Heckman, and Susanna Miettinen
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melflufen ,melphalan flufenamide ,peptide–drug conjugate ,mesenchymal stem/stromal cells ,multiple myeloma ,bone marrow ,Cytology ,QH573-671 - Abstract
Mesenchymal stem/stromal cells (MSCs) are self-renewing and multipotent progenitors, which constitute the main cellular compartment of the bone marrow stroma. Because MSCs have an important role in the pathogenesis of multiple myeloma, it is essential to know if novel drugs target MSCs. Melflufen is a novel anticancer peptide–drug conjugate compound for patients with relapsed refractory multiple myeloma. Here, we studied the cytotoxicity of melflufen, melphalan and doxorubicin in healthy human bone marrow-derived MSCs (BMSCs) and how these drugs affect BMSC proliferation. We established co-cultures of BMSCs with MM.1S myeloma cells to see if BMSCs increase or decrease the cytotoxicity of melflufen, melphalan, bortezomib and doxorubicin. We evaluated how the drugs affect BMSC differentiation into adipocytes and osteoblasts and the BMSC-supported formation of vascular networks. Our results showed that BMSCs were more sensitive to melflufen than to melphalan. The cytotoxicity of melflufen in myeloma cells was not affected by the co-culture with BMSCs, as was the case for melphalan, bortezomib and doxorubicin. Adipogenesis, osteogenesis and BMSC-mediated angiogenesis were all affected by melflufen. Melphalan and doxorubicin affected BMSC differentiation in similar ways. The effects on adipogenesis and osteogenesis were not solely because of effects on proliferation, seen from the differential expression of differentiation markers normalized by cell number. Overall, our results indicate that melflufen has a significant impact on BMSCs, which could possibly affect therapy outcome.
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- 2022
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4. Prognostic significance of esterase gene expression in multiple myeloma
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Muntasir Mamun Majumder, Romika Kumari, Nina N. Nupponen, Raija Silvennoinen, Pekka Anttila, Juha Lievonen, Fredrik Lehmann, Caroline A. Heckman, Institute for Molecular Medicine Finland, Helsinki Institute of Life Science HiLIFE, Digital Precision Cancer Medicine (iCAN), Hematologian yksikkö, HUS Comprehensive Cancer Center, Department of Oncology, and University of Helsinki
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Adult ,Male ,Oncology ,Cancer Research ,medicine.medical_specialty ,Esterase Gene ,3122 Cancers ,Esterase ,Article ,Cohort Studies ,Tumour biomarkers ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Exome Sequencing ,medicine ,Humans ,Finland ,Multiple myeloma ,Aged ,030304 developmental biology ,Aged, 80 and over ,0303 health sciences ,Sequence Analysis, RNA ,business.industry ,Gene Expression Profiling ,Esterases ,High-Throughput Nucleotide Sequencing ,Cancer ,Middle Aged ,Prognosis ,medicine.disease ,PON1 ,Molecular medicine ,3. Good health ,Gene Expression Regulation, Neoplastic ,Gene expression profiling ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Female ,Bone marrow ,Multiple Myeloma ,business - Abstract
Background Esterase enzymes differ in substrate specificity and biological function and may display dysregulated expression in cancer. This study evaluated the biological significance of esterase expression in multiple myeloma (MM). Methods For gene expression profiling and evaluation of genomic variants in the Institute for Molecular Medicine Finland (FIMM) cohort, bone marrow aspirates were obtained from patients with newly diagnosed MM (NDMM) or relapsed/refractory MM (RRMM). CD138+ plasma cells were enriched and used for RNA sequencing and analysis, and to evaluate genomic variation. The Multiple Myeloma Research Foundation (MMRF) Relating Clinical Outcomes in MM to Personal Assessment of Genetic Profile (CoMMpass) dataset was used for validation of the findings from FIMM. Results MM patients (NDMM, n = 56; RRMM, n = 78) provided 171 bone marrow aspirates (NDMM, n = 56; RRMM, n = 115). Specific esterases exhibited relatively high or low expression in MM, and expression of specific esterases (UCHL5, SIAE, ESD, PAFAH1B3, PNPLA4 and PON1) was significantly altered on progression from NDMM to RRMM. High expression of OVCA2, PAFAH1B3, SIAE and USP4, and low expression of PCED1B, were identified as poor prognostic markers (P PAFAH1B3 and SIAE, and lower expression of PCED1B, were associated with poor prognosis. Conclusions Esterase gene expression levels change as patients progress from NDMM to RRMM. High expression of OVCA2, PAFAH1B3, USP4 and SIAE, and low expression of PCED1B, are poor prognostic markers in MM, suggesting a role for these esterases in myeloma biology.
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- 2021
5. Single Cell RNA Sequencing Identifies Potential Molecular Indicators of Response to Melflufen in Multiple Myeloma
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Ana Slipicevic, Caroline A. Heckman, Sadiksha Adhikari, Nina N. Nupponen, Philipp Sergeev, Minna Suvela, Fredrik Lehmann, Maiju-Emilia Huppunen, and Juho J. Miettinen
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0303 health sciences ,Immunology ,Cell ,RNA ,Cell Biology ,Hematology ,Computational biology ,Biology ,medicine.disease ,Biochemistry ,3. Good health ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,medicine ,Multiple myeloma ,030304 developmental biology ,030215 immunology - Abstract
Introduction Melphalan flufenamide (melflufen), is a novel peptide-drug conjugate that targets aminopeptidases and selectively delivers alkylating agents in tumors. Melflufen was recently FDA approved for the treatment of relapsed/refractory multiple myeloma (MM) patients. Considering the challenges in treating this group of patients, and the availability of several new drugs for MM, information that can support treatment selection is urgently needed. To identify potential indicators of response and mechanism of resistance to melflufen, we applied a multiparametric drug sensitivity assay to MM patient samples ex vivo and analyzed the samples by single cell RNA sequencing (scRNAseq). Ex vivo drug testing identified MM samples that were distinctly sensitive or resistant to melflufen, while differential gene expression analysis revealed pathways associated with response. Methods Bone marrow (BM) aspirates from 24 MM patients were obtained after written informed consent following approved protocols in compliance with the Declaration of Helsinki. BM mononuclear cells from 12 newly diagnosed (ND) and 12 relapsed/refractory (RR) patients were used for multi-parametric flow cytometry-based drug sensitivity and resistance testing (DSRT) evaluation to melflufen and melphalan, and for scRNAseq. Based on the results from the DSRT tests and drug sensitivity scores (DSS), we divided the samples into three groups - high sensitivity (HS, DSS > 40 (melflufen) or DSS > 16 (melphalan)), intermediate sensitivity (IS, 31 ≤ DSS ≤ 40 (melflufen) or 10 ≤ DSS ≤ 16 (melphalan)), and low sensitivity (LS, DSS < 31 (melflufen) or DSS < 10 (melphalan)). To identify genes, responsible for the general sensitivity to melphalan-based drugs we conducted differential gene expression (DGE) analyses separately for melphalan and melflufen focusing on the plasma cell populations, comparing gene expression between HS and LS samples for both drugs ("HS vs. LS melphalan" and "HS vs. LS for melflufen", respectively). In addition, to explain the increased sensitivity of RR samples, we conducted the DGE analysis for ND vs. RR samples and searched for similarities between these three datasets. Results DSRT data indicated that samples from RRMM patients were significantly more sensitive to melflufen compared to samples from NDMM (Fig. 1A). In addition, we observed that samples with a gain of 1q (+1q) were more sensitive to melflufen while those with deletion of 13q (del13q) appeared to be less sensitive, although these results lacked significance (Fig. 1A). After separating the samples into different drug sensitivity groups (HS, IS, LS), DGE analysis showed significant downregulation of the drug efflux and multidrug resistance protein family member ABCB9 in the melflufen HS group opposed to the LS group (2.2-fold, p < 0.001). A similar pattern was detected for the melphalan HS vs. LS comparison suggesting that this alteration might be a common indicator of sensitivity to melphalan-based drugs. Furthermore, in the melflufen HS group we observed downregulation of the matrix metallopeptidase inhibitors TIMP1 and TIMP2 (3-fold and 1.6-fold, p < 0.001, respectively), and cathepsin inhibitors CST3 and CSTB (3.2-fold and 1.3-fold, p < 0.001, respectively) (Fig. 1B). This effect was observed in both "ND vs. RR" and "HS vs. LS for melflufen" comparisons, but not for melphalan, suggesting that these changes are associated with disease progression and specific indicators of sensitivity to melflufen. Moreover, gene set enrichment analysis (GSEA) showed activation of pathways related to protein synthesis, as well as amino acid starvation for malignant and normal cell populations in the HS group. Conclusion In summary, our results indicate that melflufen is more active in RRMM compared to NDMM. In addition, samples from MM patients with +1q, which is considered an indicator of high-risk disease, tended to be more sensitive to melflufen. Based on differential GSEA and pathway enrichment, several synergizing mechanisms could potentially explain the higher sensitivity to melflufen, such as decreased drug efflux and increased drug uptake. Although these results indicate potential indicators of response and mechanisms of drug efficacy, further validation of these findings is required using data from melflufen treated patients. Figure 1 Figure 1. Disclosures Slipicevic: Oncopeptides AB: Current Employment. Nupponen: Oncopeptides AB: Consultancy. Lehmann: Oncopeptides AB: Current Employment. Heckman: Orion Pharma: Research Funding; Oncopeptides: Consultancy, Research Funding; Novartis: Research Funding; Celgene/BMS: Research Funding; Kronos Bio, Inc.: Research Funding.
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- 2021
6. Combination of Melphalan Flufenamide and Anti-PD-L1 or Anti-CD38 Antibodies Enhances Anti-Myeloma Immunity
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Nina N. Nupponen, Kenneth C. Anderson, Paul G. Richardson, Joachim Gullbo, Dharminder Chauhan, Fredrik Lehmann, Arghya Ray, Ting Du, and Jakob Lindberg
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biology ,business.industry ,Immunology ,Anti pd 1 ,Cell Biology ,Hematology ,CD38 ,Biochemistry ,Melphalan-flufenamide ,Immunity ,Cancer research ,biology.protein ,Medicine ,Antibody ,business - Abstract
Introduction Melphalan flufenamide (Melflufen; Oncopeptides AB) is a novel enzyme-activated analogue of melphalan that enables a more rapid and higher intracellular accumulation of melphalan in tumor cells than is achievable by direct exposure to equimolar doses of melphalan. Our preclinical study showed that melflufen is a more potent anti-myeloma (MM) agent than melphalan, overcomes drug-resistance, and induces synergistic anti-MM activity in combination with bortezomib, lenalidomide, or dexamethasone (Chauhan et al, Clinical Cancer Res 2013;19:3019). However, the effect of melflufen on the immunosuppressive and tumor-promoting MM-host bone marrow (BM) accessory cells such as immunologically dysfunctional plasmacytoid dendritic cells (pDCs; CD123/IL-3Rα) remains unclear. Here, we utilized our coculture models of pDCs, T-, and NK cells with autologous patient MM cells to examine whether a combination of melflufen and immune checkpoint inhibitor anti-PD-L1 Ab, or daratumumab (anti-CD38 Ab), restores anti-MM immunity. Methods MM patient BM and PB samples (N=10; obtained after informed consent), and cell lines were used for the study. Minimally cytotoxic concentration of melflufen (0.1 µM) was used to assess immune functions. CTL/NK activity assays MM CD8 + T- or NK-cells were cultured with autologous pDCs (1:10 pDC:T/NK ratio) with melflufen (0.1 μM) alone, and with anti-PD-L1 (5 μg/ml) or anti-CD38 (0.5 μg/ml) Abs for 3-5 days; cells were washed to remove the drugs, and then cultured for another 24h with pre-stained target MM cells (10:1 E/T ratio; T/NK:MM), followed by quantification of viable MM cells by flow. Results 1) Both MM tumor cells and pDCs showed higher PD-L1 and CD38 levels vs normal plasma cells; 2) Treatment of MM patient total BM mononuclear cells or purified MM cells with melflufen (0.1 µM) increased PD-L1 expression on MM cells (1.84-fold, treated vs untreated; p Conclusions The combination of melflufen and anti-PD-L1 increases pDC-induced T- and NK cell-mediated cytolytic activities against MM. Moreover, combined melflufen and anti-CD38 Abs modestly enhance pDC-induced NK cell-mediated MM-specific cytolytic activity. Our preclinical data suggest targeting PD-L1 in combination with melflufen as well as support an ongoing clinical trial of melflufen with anti-CD38 Abs to enhance anti-MM immunity. Disclosures Nupponen: Oncopeptides AB: Consultancy. Lehmann: Oncopeptides AB: Current Employment. Lindberg: Oncopeptides: Current Employment, Current equity holder in publicly-traded company, Divested equity in a private or publicly-traded company in the past 24 months, Other: Travel, Accommodations, Expenses; Camurus: Membership on an entity's Board of Directors or advisory committees, Other: Travel, Accommodations, Expenses; Affibody: Membership on an entity's Board of Directors or advisory committees. Gullbo: Oncopeptides AB: Consultancy. Richardson: Takeda: Consultancy, Research Funding; Celgene/BMS: Consultancy, Research Funding; Janssen: Consultancy; Sanofi: Consultancy; Protocol Intelligence: Consultancy; Karyopharm: Consultancy, Research Funding; GlaxoSmithKline: Consultancy; Regeneron: Consultancy; AstraZeneca: Consultancy; Secura Bio: Consultancy; AbbVie: Consultancy; Oncopeptides: Consultancy, Research Funding; Jazz Pharmaceuticals: Consultancy, Research Funding. Chauhan: C4 Therapeutics: Current equity holder in publicly-traded company; Oncopeptides: Consultancy; Stemline Therapeutics: Consultancy. Anderson: Janssen: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees; Sanofi-Aventis: Membership on an entity's Board of Directors or advisory committees; Bristol Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Millenium-Takeda: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Scientific Founder of Oncopep and C4 Therapeutics: Current equity holder in publicly-traded company, Current holder of individual stocks in a privately-held company; AstraZeneca: Membership on an entity's Board of Directors or advisory committees; Mana Therapeutics: Membership on an entity's Board of Directors or advisory committees.
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- 2021
7. Abstract 1843: Melflufen efficacy in multiple myeloma with TP53 aberrations
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Caroline A. Heckman, Thorsten Stühmer, Juho J. Miettinen, Maria-Victoria Mateos, Johan Aschan, Paul G. Richardson, Nina N. Nupponen, Ralf C. Bargou, Paula Rodriguez Otero, Maiju-Emilia Huppunen, Fredrik Lehmann, Ana Slipicevic, and Umair Munawar
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0301 basic medicine ,Melphalan ,Cancer Research ,medicine.diagnostic_test ,business.industry ,Cancer ,Phases of clinical research ,Plasma cell ,medicine.disease ,3. Good health ,Flow cytometry ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,Oncology ,Apoptosis ,030220 oncology & carcinogenesis ,medicine ,Cancer research ,business ,Ex vivo ,Multiple myeloma ,medicine.drug - Abstract
Introduction Multiple myeloma (MM) is an incurable plasma cell malignancy characterized by clonal evolution and heterogeneous genetic abnormalities. Deletion of the short arm of chromosome 17 (del17p) harboring TP53 is a high-risk abnormality associated with aggressive disease and therapy resistance. Here, we tested in vitro and ex vivo efficacy of a peptide-drug conjugate melflufen, currently in phase 3 clinical trials for relapsed/refractory multiple myeloma (RRMM), in an isogenic myeloma p53-abrogated cell line model and patient samples. Methods Efficacy of melflufen versus melphalan was tested in the AMO-1 cell line, which displays biallelic wildtype TP53 and retains aspects of a functional p53 system, and AMO-1 clones with either complete loss of p53 or expressing point-mutated p53 protein (R282W hotspot mutation), created using the CRISPR/Cas9 system and modified via Sleeping Beauty vectors. We assessed toxicity and apoptosis using Annexin V and Alamar blue assays. Ex vivo sensitivity to the drugs was examined by flow cytometry in CD138+ plasma cells isolated from MM patients with confirmed del17p or TP53 mutations. We also analyzed response rates in a subpopulation of patients with confirmed del17p from OP-106 HORIZON, an ongoing phase 2 study evaluating the efficacy of melflufen plus dexamethasone in patients with RRMM (NCT02963493). Results Whereas loss of p53 functionality strongly impaired melphalan induced cytotoxicity in the AMO-1 MM cell line model system, treatment with melflufen showed superior efficacy in the parental cells, and was effective independent of the respective TP53 status. Although at low dosages wild-type cells were still more sensitive, in contrast to melphalan this effect was overcome by slight dose increases of melflufen, due to the much steeper dose-effect relationships in the p53 deficient sublines. In contrast to melphalan, the melflufen effect was only partially blocked by combined pre-treatment with inhibitors of apoptosis and necroptosis. Similar results were observed in patient-derived material with melflufen displaying superior activity towards TP53 mutated plasma cells. Analysis of OP-106 HORIZON trial data showed comparable ORR of 20% (CI 95% 4.3-48.1) in the del17p subpopulation (n=15) to 26.2% (CI 95% 18.8-34.6) in the total population (n=136). Conclusions Melflufen is able to trigger myeloma cell death regardless of p53 status, and thus overcome p53-deficiency-mediated melphalan resistance. It demonstrates better ex vivo efficacy in MM patient derived plasma cells harboring del17p or TP53 mutations. Melflufen response rates in the del17p Citation Format: Ana Slipicevic, Umair Munawar, Thorsten Stühmer, Johan Aschan, Fredrik Lehmann, Nina N. Nupponen, Juho Miettinen, Maiju-Emilia Huppunen, Caroline A. Heckman, María-Victoria Mateos, Paula Rodríguez Otero, Paul Richardson, Ralf Bargou. Melflufen efficacy in multiple myeloma with TP53 aberrations [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1843.
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- 2020
8. Abstract 190: Prognostic significance of esterase gene expression in multiple myeloma
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Raija Silvennoinen, Caroline A. Heckman, Pekka Anttila, Juha Lievonen, Romika Kumari, Fredrik Lehmann, Mamun Majumder, and Nina N. Nupponen
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Cancer Research ,Oncology ,Esterase Gene ,Cancer research ,medicine ,Biology ,medicine.disease ,Multiple myeloma - Abstract
Introduction: The esterase enzyme family is a subclass of the hydrolase enzyme superfamily that functions to split ester bonds. Several esterases have been identified, which differ in substrate specificity and biological function, and the expression of specific esterases may be dysregulated in cancer. Limited information is available regarding the expression of esterases in multiple myeloma (MM), despite potential for exploitation in novel therapeutic approaches such as peptide-drug conjugates (e.g. melflufen). We evaluated the biological significance of esterase enzyme expression in MM. Methods: For gene expression profiling, bone marrow aspirates were obtained from MM patients (pts) or healthy donors after obtaining written informed consent and following approved protocols in compliance with the Declaration of Helsinki. CD138+ plasma cells were enriched and used for RNA preparation. Illumina-compatible RNA sequencing libraries were prepared and sequenced. A cut-off value of >1 Reads Per Kilobase of transcript per Million mapped reads (RPKM) was used to filter expressed genes. The Welch two-sample t-test was used to identify significant variation in gene expression. Contribution of esterase gene expression to survival outcome was estimated by Kaplan-Meier analysis; significance between two groups was deduced using a Mantel-Cox log rank test. Results: Of 123 pts with MM, bone marrow aspirates were collected from 41 pts with newly diagnosed (ND) MM, 81 pts with relapsed/refractory (RR) MM, and 1 pt with stable disease. Samples were also obtained from two healthy donors. The most abundant esterases in MM samples were OVCA2, PAFAH1B2, NXPE3, UCHL3, LIPA, ABHD10, UCHL5, CASD1, ABHD13, and USP4 (median log2[RPKM] range: 3.9 to 2.1). The least abundant esterases were NLGN1, NLGN2, PON1, CEL, PON3, NLGN4Y, IL17A, AADAC, CES5A, and ASPG (median log2[RPKM]: -5.1 to -10.7). The expression levels of OVCA2, PAFAH1B2, NXPE3, UCHL5, SIAE, ESD, PNPLA6, and PAFAH1B3 were significantly higher (P Conclusion: Esterase expression levels in MM change on progression from NDMM to RRMM. High expression of OVCA2, PAFAH1B3, CES4A, and PNPLA6, and low expression of NXPE4 and GZMA, were identified as poor prognostic markers in pts with MM, suggesting a role for these esterases in myeloma biology. Citation Format: Romika Kumari, M. Mamun Majumder, Juha Lievonen, Raija Silvennoinen, Pekka Anttila, Nina N. Nupponen, Fredrik Lehmann, Caroline A. Heckman. Prognostic significance of esterase gene expression in multiple myeloma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 190.
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- 2020
9. In Vitro and inVivo Activity of Melflufen in Amyloidosis
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Caroline A. Heckman, Minna Suvela, Muntasir Mamun Majumder, Fredrik Lehmann, Sarah A. Holstein, Ana Slipicevic, Ken Flanagan, Juho J. Miettinen, Romika Kumari, Michelle L. Varney, and Nina N. Nupponen
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0301 basic medicine ,Melphalan ,Immunology ,Plasma cell ,Immunoglobulin light chain ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,medicine ,AL amyloidosis ,Multiple myeloma ,biology ,Chemistry ,Amyloidosis ,Cell Biology ,Hematology ,medicine.disease ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,biology.protein ,Cancer research ,Bone marrow ,Antibody ,030215 immunology ,medicine.drug - Abstract
Background: Immunoglobulin light-chain (AL) amyloidosis is a rare disease caused by plasma cell secretion of misfolded light chains that assemble as amyloid fibrils and deposit on vital organs including the heart and kidneys, causing organ dysfunction. Plasma cell directed therapeutics, aimed at preferentially eliminating the clonal population of amyloidogenic cells in bone marrow are expected to reduce production of toxic light chain and alleviate deposition of amyloid thereby restoring healthy organ function. Melphalan flufenamide ethyl ester, melflufen, is a peptidase potentiated alkylating agent with potent toxicity in myeloma cells. Melflufen is highly lipophilic, permitting rapid cellular uptake, and is subsequently enzymatically cleaved by aminopeptidases within cells resulting in augmented intracellular concentrations of toxic molecules, providing a more targeted and localized treatment. Previous data demonstrating multiple myeloma plasma cell sensitivity for melflufen suggests that the drug might be useful to directly eliminate amyloidogenic plasma cells, thereby reducing the amyloid load in patients. Furthermore, the increased intracellular concentrations of melflufen in myeloma cells indicates a potential reduction in systemic toxicity in patients, an important factor in the fragile amyloidosis patient population. To assess potential efficacy in amyloidosis patients and to explore the mechanism of action, we examined effects of melflufen on amyloidogenic plasma cells invitro and invivo. Methods: Cellular toxicity and apoptosis were measured in response to either melflufen or melphalan in multiple malignant human plasma cell lines, including the amyloidosis patient derived light chain secreting ALMC-1 and ALMC-2 cells, as well as primary bone marrow cells from AL amyloidosis patients, using annexin V and live/dead cell staining by multicolor flow cytometry, and measurement of cleaved caspases. Lambda light chain was measured in supernatant by ELISA, and intracellular levels were detected by flow cytometry. To assess efficacy of melflufen in vivo, the light chain secreting human myeloma cell line, JJN3, was transduced with luciferase and adoptively transferred into NSG mice. Cell death in response to melflufen or melphalan was measured by in vivo bioluminescence, and serum light chain was monitored. Results: Melflufen demonstrated increased potency against multiple myeloma cell lines compared to melphalan, inducing malignant plasma cell death at lower doses on established light chain secreting plasma cell lines. While ALMC-1 cells were sensitive to both melphalan and melflufen, the IC50 for melphalan at 960 nM was approximately 3-fold higher than melflufen (334 nM). However, ALMC-2 cells were relatively insensitive to melphalan (12600 nM), but maintained a 100-fold increase in sensitivity to melflufen (121 nM). Furthermore, while 40% of primary CD138+ plasma cells from patients with diagnosed AL amyloidosis responded to melflufen treatment in vitro, only 20% responded to melphalan with consistently superior IC50 values for melflufen (Figure 1). Light chain secreting cell lines and AL amyloidosis patient samples were further analyzed by single cell sequencing. We further examined differential effects on apoptosis and the unfolded protein response in vitro in response to either melflufen or melphalan. This is of particular interest in amyloidosis, where malignant antibody producing plasma cells possess an increased requirement for mechanisms to cope with the amplified load of unfolded protein and associated ER stress. As AL amyloidosis is ultimately a disease mediated by secretion of toxic immunoglobulin, we assessed the effects of melflufen on the production of light chain invitro, measuring a decrease in production of light chain in response to melflufen treatment. Finally, we took advantage of a recently described adoptive transfer mouse model of amyloidosis to assess the efficacy of melflufen and melphalan in eliminating amyloidogenic clones and reducing the levels of toxic serum light chain in vivo. Conclusions: These findings provide evidence that melflufen mediated toxicity, previously described in myeloma cells, extends to amyloidogenic plasma cells and further affects the ability of these cells to produce and secrete toxic light chain. This data supports the rationale for the evaluation of melflufen in patients with AL amyloidosis. Figure 1 Disclosures Flanagan: Oncopeptides AB: Employment. Slipicevic:Oncopeptides AB: Employment. Holstein:Celgene: Consultancy; Takeda: Membership on an entity's Board of Directors or advisory committees; Adaptive Biotechnologies: Membership on an entity's Board of Directors or advisory committees; GSK: Consultancy; Genentech: Membership on an entity's Board of Directors or advisory committees; Sorrento: Consultancy. Lehmann:Oncopeptides AB: Employment. Nupponen:Oncopeptides AB: Employment. Heckman:Celgene: Research Funding; Novartis: Research Funding; Oncopeptides: Research Funding; Orion Pharma: Research Funding.
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- 2019
10. A phase II trial of gefitinib in patients with rising PSA following radical prostatectomy or radiotherapy
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Mirja Ruutu, Nina N. Nupponen, Greetta Joensuu, Akseli Hemminki, Timo Joensuu, Sari Pesonen, and Juhani Collan
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Oncology ,medicine.medical_specialty ,medicine.medical_treatment ,030232 urology & nephrology ,Urology ,Context (language use) ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,Gefitinib ,Prostate ,Internal medicine ,medicine ,Radiology, Nuclear Medicine and imaging ,In patient ,Epidermal growth factor receptor ,biology ,Prostatectomy ,business.industry ,Hematology ,General Medicine ,medicine.disease ,3. Good health ,Radiation therapy ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,biology.protein ,business ,medicine.drug - Abstract
Prostate cancer recurrence after therapy with curative intent is often first detected as an increase in prostate-specific antigen (PsA) (“rising PsA syndrome”) which may occur years in advance of clinical recurrence, and may present on opportunity for effective treatment in the context of minimal disease load. Eventually 5–66% of patients with rising PsA receive salvage therapies, the most common of which are radiation of the prostate basin after surgery and hormonal treatments especially in relapses following radical radiotherapy of prostate [1,2]. regrettably, although both modalities are effec-tive in many patients, they also cause significant mor-bidity. Also, radiation is effective only in locally recurrent disease and therefore only 55–72% of rising PsA patients feature PsA response when given salvage radi -ation [3,4]. Hormonal therapies are initially effective in most patients, but they cause numerous long-term side effects and their benefit is often eventually lost, which results in difficult-to-treat castration resistant prostate cancer (CrPC) [5].High expression of epidermal growth factor receptor (
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- 2011
11. Amplification and overexpression of KIT, PDGFRA, and VEGFR2 in medulloblastomas and primitive neuroectodermal tumors
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Olli Tynninen, Hannu Haapasalo, Tea Blom, Kristiina Nordfors, Miikka Korja, Nina N. Nupponen, Valtteri Häyry, Kirmo Wartiovaara, and Annariikka Roselli
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Male ,Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Gene Dosage ,Kaplan-Meier Estimate ,PDGFRA ,Biology ,Gene dosage ,Receptor tyrosine kinase ,Growth factor receptor ,medicine ,Humans ,Neuroectodermal Tumors, Primitive ,Cerebellar Neoplasms ,CISH ,In Situ Hybridization ,Medulloblastoma ,Stem Cell Factor ,Brain Neoplasms ,Kinase insert domain receptor ,medicine.disease ,Immunohistochemistry ,Vascular Endothelial Growth Factor Receptor-2 ,digestive system diseases ,Neurology ,Oncology ,Cancer research ,biology.protein ,Neurology (clinical) - Abstract
Medulloblastomas (MB) and primitive neuroectodermal tumors (PNET) are the most common malignant brain tumors in children. These two tumor types are histologically similar, but have different genetic backgrounds and clinical outcomes. Other brain tumors, such as gliomas, frequently have coamplification and overexpression of receptor tyrosine kinases KIT, platelet-derived growth factor receptor alpha (PDGFRA), and vascular endothelial growth factor receptor 2 (VEGFR2). We investigated protein expression and gene copy numbers of KIT, PDGFRA, and VEGFR2 in 41 MB and 11 PNET samples by immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH). KIT and PDGFRA expression was detected in both MBs and PNETs, whereas VEGFR2 expression was weak in these tumors. KIT, PDGFRA, and VEGFR2 amplifications were all present in 4% of MBs/PNETs, and KIT amplification was associated with concurrent PDGFRA and VEGFR2 amplifications (Por= 0.001). Most strikingly, increased gene copy number of PDGFRA was associated with poor overall survival (P = 0.027). We suggest that coamplification of PDGFRA or VEGFR2 with KIT may be clinically useful novel molecular markers in MBs and PNETs.
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- 2009
12. Mutation and copy number analysis of LNX1 and Numbl in nervous system tumors
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Tea Blom, Minna Tanner, Nina N. Nupponen, and Annariikka Roselli
- Subjects
Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Ubiquitin-Protein Ligases ,Nervous System Neoplasms ,Gene Dosage ,Copy number analysis ,Chromogenic in situ hybridization ,Locus (genetics) ,PDGFRA ,Biology ,Polymorphism, Single Nucleotide ,03 medical and health sciences ,Exon ,0302 clinical medicine ,Gene duplication ,Genetics ,Humans ,Molecular Biology ,Gene ,In Situ Hybridization ,030304 developmental biology ,0303 health sciences ,Intracellular Signaling Peptides and Proteins ,Exons ,Glioma ,Molecular biology ,Proto-Oncogene Proteins c-kit ,Mutation ,NUMB ,030217 neurology & neurosurgery - Abstract
Alterations at chromosome locus 4q12 are frequently found in gliomas; this locus contains the receptor tyrosine kinase–encoding genes KIT , PDGFRA , and KDR (alias VEGFR2 ). Notable among the genes at this locus is LNX1 , the ligand of Numb protein X. LNX1 encodes a PDZ domain containing protein, which interacts with the cell fate determinant Numbl, a Numb homolog-like gene involved in the maintenance of neural progenitor cells during embryonic neurogenesis. We performed a mutation analysis for LNX1 and Numbl genes. In addition, gene copy numbers of LNX1 , Numbl , and KIT in human nervous system tumors were analyzed by chromogenic in situ hybridization. Tissue samples from 90 patients were screened for LNX1 and Numbl mutations, and tissue sections from 56 samples were analyzed for gene amplification status. Our analysis revealed missense mutations in LNX1 exons 3 and 5 and a single-nucleotide polymorphism in Numbl exon 6. In addition, polyglutamine repeat polymorphism was found in Numbl exon 10. Chromogenic in situ hybridization showed gene amplification of LNX1 in 10%, Numbl in 5%, and KIT in 6% of nervous system tumors. Both gene sequence alterations and amplifications of LNX1 and Numbl are present in a subset of human gliomas, and the role of these genes in neurogenesis suggests that they may contribute to development of glial tumors.
- Published
- 2008
13. Stem cell protein BMI-1 is an independent marker for poor prognosis in oligodendroglial tumours
- Author
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Hannu Sariola, Anders Paetau, Valtteri Häyry, Johannes Wölfer, Olli Tynninen, Seppo Sarna, Nina N. Nupponen, Hannu Haapasalo, Kirmo Wartiovaara, Werner Paulus, Martin Hasselblatt, and Mika Niemelä
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Adult ,Male ,Pathology ,medicine.medical_specialty ,Histology ,Mitotic index ,Adolescent ,Proliferative index ,Gene Expression ,Kaplan-Meier Estimate ,Biology ,medicine.disease_cause ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Proto-Oncogene Proteins ,Physiology (medical) ,Glioma ,Biomarkers, Tumor ,medicine ,Humans ,Cyclin-Dependent Kinase Inhibitor p16 ,Aged ,030304 developmental biology ,Aged, 80 and over ,Polycomb Repressive Complex 1 ,0303 health sciences ,Brain Neoplasms ,Nuclear Proteins ,Cancer ,Proto-Oncogene Proteins c-mdm2 ,Middle Aged ,medicine.disease ,Immunohistochemistry ,3. Good health ,Repressor Proteins ,Neurology ,030220 oncology & carcinogenesis ,Female ,Neurology (clinical) ,Oligodendroglioma ,Stem cell ,Carcinogenesis - Abstract
Aims: The polycomb factor BMI-1 has recently been implicated in tumorigenesis of the central nervous system in several experimental animal models. However, the significance of BMI-1 in human glioma has not been investigated. Here we describe expression of the polycomb protein BMI-1 and its downstream targets p16Ink4a and MDM2 in both high- and low-grade human glioma. Methods: Tumour samples were collected from 305 adult patients treated for primary grades 2–4 gliomas between 1980 and 2006 in Finland and Germany. BMI-1, p16 and MDM2 expression was evaluated using immunohistochemistry in representative paraffin-embedded tumour tissue. The significance of observed immunoreactivity, age at onset, gender, histopathological findings and proliferative index was analysed in univariate and multivariate survival models. Results: BMI-1 was expressed in all histologic types of diffuse gliomas. We found a significant correlation (P = 0.007) between the frequency of BMI-1 immunoreactive tumour cells and poor survival in World Health Organization grades II–III oligodendrogliomas and oligoastrocytomas (n = 62). The median survival of patients grouped by low, intermediate or high frequency of BMI-1 immunoreactive tumour cells was 191 months, 151 months and 68 months, respectively. This association was also significant in the Cox multivariate regression model. Nuclear p16 immunopositivity predicted better survival in astrocytomas and an inverse correlation between p16 expression and the Ki-67 mitotic index was also observed. Conclusions: BMI-1 is found in all histological types of gliomas and the relative protein expression of BMI-1 is a novel independent prognostic marker in oligodendroglial tumours.
- Published
- 2008
14. KIT overexpression induces proliferation in astrocytes in an imatinib-responsive manner and associates with proliferation index in gliomas
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Nina N. Nupponen, Kirmo Wartiovaara, Tea Blom, Heli Fox, Hannu Haapasalo, Alexandre Angers-Loustau, Laura Kerosuo, Olli A. Jänne, Karita Peltonen, Panu E. Kovanen, and Marika J. Linja
- Subjects
0303 health sciences ,Cancer Research ,Proliferation index ,medicine.drug_class ,Cell growth ,Biology ,Gene mutation ,Tyrosine-kinase inhibitor ,3. Good health ,03 medical and health sciences ,0302 clinical medicine ,Imatinib mesylate ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,medicine ,Proto-Oncogene Proteins c-kit ,Receptor Tyrosine Kinase Gene ,Signal transduction ,030304 developmental biology - Abstract
Activating gene mutations, gene amplifications and overexpressed proteins may be useful as targets for novel therapies. Alterations at chromosome locus 4q12 are associated with gliomas and the region harbors the receptor tyrosine kinase gene KIT, which is frequently amplified in gliomas, and also overexpressed in a subset of gliomas. KIT and its ligand stem cell factor are widely expressed in embryonic and adult mouse brain, and they play a role in many signal transduction pathways involved in cellular proliferation, differentiation and cancer cell metastasis. However, the function of KIT in gliomagenesis or disease progression remains unresolved as well as its role in neural and brain tumor development. In this study, we utilized lentivirus-mediated gene transfer to deliver the KIT gene into mouse astrocytes. The growth properties of KIT overexpressing cells were analyzed using several in vitro functional assays. The effect of receptor tyrosine kinase inhibitor imatinib on astrocyte growth was also investigated. Our results indicate that overexpression of KIT in mouse astrocytes promotes cell proliferation, and the increased proliferation is partly inhibited by imatinib treatment. Furthermore, KIT overexpression induces phenotypic changes in the cells suggesting that KIT may play a role in astrocyte growth regulation.
- Published
- 2008
15. Copy number alterations of the polycomb gene BMI1 in gliomas
- Author
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Olli Tynninen, Miina Ollikainen, Minna Tanner, Tea Blom, Annariikka Roselli, Valtteri Häyry, Hannu Sariola, Nina N. Nupponen, and Kirmo Wartiovaara
- Subjects
Adult ,Male ,Adolescent ,Gene Dosage ,Locus (genetics) ,Kaplan-Meier Estimate ,macromolecular substances ,Polymerase Chain Reaction ,Pathology and Forensic Medicine ,Cellular and Molecular Neuroscience ,Proto-Oncogene Proteins ,Glioma ,Centromere ,medicine ,Humans ,CISH ,Gene ,In Situ Hybridization ,Aged ,Polycomb Repressive Complex 1 ,biology ,Brain Neoplasms ,Nuclear Proteins ,Middle Aged ,Prognosis ,medicine.disease ,Molecular biology ,Neural stem cell ,Repressor Proteins ,Histone ,BMI1 ,biology.protein ,Female ,Neurology (clinical) - Abstract
Gliomas are heterogeneous tumours that grow in an uninhibited fashion, and these brain tumour cells share numerous characteristics with neural stem cells. The BMI1 gene encodes a component of the polycomb protein complex regulating epigenetically gene activity via histone modiWcation. It functions for instance during the develop- ment of the central nervous system and maturation of neu- ral cells. BMI-1 protein expression is deregulated in several forms of cancer and gene ampliWcation has been identiWed in mantle cell lymphomas. Since BMI1 is located at chro- mosome 10p, a region implicated frequently in brain tumo- urigenesis, we investigated the genetic status and the corresponding expression patterns of BMI1 in a series of 100 low- and high-grade primary and recurrent gliomas. Chromogenic in situ hybridisation (CISH) with probes directed against BMI1 at 10p13 and the centromere of chro- mosome 10 was used in the analyses. Of all gliomas, 59% demonstrated aberrant copy numbers of BMI1. Deletions of the BMI1 locus were found in most types of tumours, and in a univariate survival analysis these cases had poor progno- sis. Increased copy numbers of the BMI1 locus (3-5 copies) were found in all histological types, especially in high- grade astrocytomas. No diVerence in prognosis between cases with normal copy numbers and cases with increased copy numbers could be observed. This data suggests that BMI1 gene is aberrant at the chromosomal level in a subset of gliomas, and possibly contributes to brain tumour patho- genesis.
- Published
- 2008
16. Platelet-derived growth factor receptor expression and amplification in choroid plexus carcinomas
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Janna Paulsson, Nina N. Nupponen, Johannes E. A. Wolff, Werner Paulus, Arne Östman, Minna Tanner, Brigitte Wrede, Martin Hasselblatt, and Astrid Jeibmann
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Male ,Choroid Plexus Neoplasms ,Pathology ,medicine.medical_specialty ,Receptor, Platelet-Derived Growth Factor alpha ,PDGFRB ,PDGFRA ,Biology ,Pathology and Forensic Medicine ,Receptor, Platelet-Derived Growth Factor beta ,03 medical and health sciences ,0302 clinical medicine ,Biomarkers, Tumor ,medicine ,Humans ,CISH ,Receptor ,030304 developmental biology ,0303 health sciences ,Carcinoma ,Gene Amplification ,Infant ,Choroid plexus carcinoma ,Prognosis ,medicine.disease ,Gene Expression Regulation, Neoplastic ,Imatinib mesylate ,Child, Preschool ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Female ,Choroid plexus ,Platelet-derived growth factor receptor - Abstract
Platelet-derived growth factor (PDGF) receptor signaling has been implicated in the development of glial tumors, but not yet been examined in choroid plexus carcinomas, pediatric tumors with dismal prognosis for which novel treatment options would be desirable. Therefore, protein expression of PDGF receptors alpha and beta as well as amplification status of the respective genes, PDGFRA and PDGFRB, were examined in a series of 22 patients harboring choroid plexus carcinoma using immunohistochemistry and chromogenic in situ hybridization (CISH). The majority of choroid plexus carcinomas expressed PDGF receptors with 6 cases (27%) displaying high staining scores for PDGF receptor alpha and 13 cases (59%) showing high staining scores for PDGF receptor beta. Correspondingly, copy-number gains of PDGFRA were observed in 8 cases out of 12 cases available for CISH and 1 case displayed amplification (six or more signals per nucleus). The proportion of choroid plexus carcinomas with amplification of PDGFRB was even higher (5/12 cases). PDGFRB amplification status and PDGF receptor beta protein expression scores were significantly correlated (P=0.01, Spearman). Expression status of PDGF receptor alpha or PDGF receptor beta was not significantly associated with progression-free survival. To conclude, expression and amplification of PDGF receptors, particularly PDGF receptor beta, are frequent in choroid plexus carcinomas, providing a first rationale for the development of treatments targeting PDGF receptor signaling in these rare malignant pediatric tumors.
- Published
- 2008
17. Molecular pathology of gliomas
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Nina N. Nupponen and Heikki Joensuu
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Pathology ,medicine.medical_specialty ,biology ,Molecular pathology ,Astrocytoma ,PDGFRA ,medicine.disease ,Receptor tyrosine kinase ,nervous system diseases ,Pathology and Forensic Medicine ,Glioma ,Chromosome 19 ,biology.protein ,medicine ,Cancer research ,PTEN ,neoplasms ,Platelet-derived growth factor receptor - Abstract
Summary Gliomas are the most common primary neoplasms of the brain. They are a heterogeneous group of tumours characterized by infiltrative growth, and relative resistance to radiotherapy and chemotherapy. Glioblastomas have complex chromosomal aberrations including amplifications and gains of 7p, 12q13–q21, and chromosome 19, and losses are 10q , 9p, 13q and 22q, whereas the karyotypes of pilocytic astrocytomas show only limited changes. Important genetic aberrations include up-regulation of EGFR and MDM2 function and loss of PTEN function in primary glioblastoma, and up-regulation of PDGFRA and CDK4 function in secondary glioblastomas that arise from a pre-existing lower grade astrocytoma. TP53, CDKNA2 and RB1 functions are often lost in secondary glioblastomas. Amplifications of PDGFRA, KIT and VEGFR2 may also have a role in the genesis of some gliomas. The proteins encoded by these genes have become targets for novel therapies, which include specific inhibitors of the EGFR, KIT, and PDGFR receptor tyrosine kinases.
- Published
- 2006
18. Molecular genetic analysis of the REST/NRSF gene in nervous system tumors
- Author
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Olli Tynninen, Maija Halonen, Hannu Haapasalo, Anders Paetau, Nina N. Nupponen, Tea Blom, Marjut Puputti, and Minna Tanner
- Subjects
Nervous System Neoplasms ,Brain tumor ,Biology ,medicine.disease_cause ,Pathology and Forensic Medicine ,Cellular and Molecular Neuroscience ,medicine ,Humans ,Copy-number variation ,CISH ,Molecular Biology ,neoplasms ,Gene ,Chromatography, High Pressure Liquid ,In Situ Hybridization, Fluorescence ,Mutation ,Chi-Square Distribution ,medicine.diagnostic_test ,Exons ,Glioma ,medicine.disease ,Molecular biology ,nervous system diseases ,Repressor Proteins ,Neurology (clinical) ,Oligodendroglioma ,Carcinogenesis ,Transcription Factors ,Fluorescence in situ hybridization - Abstract
The gene for RE1-silencing transcription factor (REST) alias neuron-restrictive silencer factor NRSF, acts as a transcriptional repressor in the neuronal differentiation pathways in non-neuronal cells, and plays an important role in neuronal development. Inactivating mutations or overexpression of REST have previously been reported in various types of cancer, but no data is available for the role of REST alterations in gliomas. REST gene was screened for mutations in 161 nervous system tumors consisting of astrocytomas, glioblastomas, oligodendrogliomas, oligoastrocytomas, medulloblastomas, meningiomas and schwannomas. REST exons 1-3 were analyzed using denaturing high-performance liquid chromatography (DHPLC) and direct sequencing. The gene copy numbers of REST were investigated by chromogenic (CISH) and fluorescence in situ hybridization (FISH) techniques. Non-synonymous SNPs (P797L, P815S) were found in eight different brain tumor samples. No truncating or activating novel mutations of REST were discovered. Since REST is located at 4q12, a chromosome region implicated in brain tumorigenesis, we conducted gene copy number analyses in medulloblastomas and gliomas. The majority of gliomas (67%) demonstrated low-level amplifications of REST, and only one oligodendroglioma showed high-level amplification of the gene. In medulloblastomas, 38% of samples were determined as aneuploidic, no high-level amplifications were found. Our data suggests that REST is neither activated nor inactivated via mutations in gliomas, while high-level amplification may rarely occur.
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- 2006
19. Gene amplification, mutation, and protein expression of EGFR and mutations of ERBB2 in serous ovarian carcinoma
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Heini Lassus, Heikki Joensuu, Jorma Isola, Nina N. Nupponen, Ralf Bützow, Harri Sihto, and Arto Leminen
- Subjects
Ovarian Neoplasms ,Proliferation index ,Receptor, ErbB-2 ,Serous carcinoma ,Gene Dosage ,Chromogenic in situ hybridization ,Middle Aged ,Biology ,medicine.disease ,Cystadenocarcinoma, Serous ,ErbB Receptors ,Serous fluid ,Ovarian carcinoma ,Mutation ,Drug Discovery ,medicine ,Cancer research ,Humans ,Molecular Medicine ,Female ,CISH ,Ovarian cancer ,In Situ Hybridization ,Genetics (clinical) ,EGFR inhibitors - Abstract
EGFR and erbB-2 are targets for specific cancer therapy. The purpose of this study was to examine the frequency and clinicopathological correlations of gene amplification, protein expression, and mutations of EGFR and ERBB2 in serous carcinoma, the most common and aggressive type of ovarian cancer. Tissue microarray constructed of 398 carcinomas was examined by chromogenic in situ hybridization (CISH) and by immunohistochemistry. Cases with amplification of EGFR by CISH were further analyzed by fluorescence in situ hybridization. One hundred ninety-eight samples were analyzed for mutations in exons 18, 19, or 21 of EGFR and in exon 20 of ERBB2 using denaturating high-performance liquid chromatography and direct sequencing. Amplification of EGFR was present in 12% (41/333), low-level gain in 43% (144/333), and protein overexpression in 17% (66/379) of the tumors. Both increased copy number and overexpression of EGFR were associated with high tumor grade, greater patient age, large residual tumor size, high proliferation index, aberrant p53, and poor patient outcome. Furthermore, increased copy number of EGFR was associated with increased copy number of ERBB2. No mutations were identified in EGFR, whereas one tumor had an insertion mutation in exon 20 of ERBB2. Both amplification and protein overexpression of EGFR occur in serous ovarian carcinoma, but EGFR copy number has a stronger prognostic value. This makes EGFR amplification a potentially useful criterion for selecting patients in clinical trials testing the effect of EGFR inhibitors in serous ovarian carcinoma.
- Published
- 2006
20. Epidermal growth factor receptor domain II, IV, and kinase domain mutations in human solid tumors
- Author
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Heikki Joensuu, Marjut Puputti, Olli Tynninen, Laura Pulli, Walter J. Koskinen, Aija Knuuttila, Minna Tanner, Nina N. Nupponen, Ralf Bützow, Harri Sihto, Tapio Visakorpi, Tom Böhling, and Leena-Maija Aaltonen
- Subjects
Lung Neoplasms ,Molecular Sequence Data ,medicine.disease_cause ,Polymerase Chain Reaction ,Receptor tyrosine kinase ,Growth factor receptor ,Carcinoma, Non-Small-Cell Lung ,Drug Discovery ,medicine ,Humans ,Amino Acid Sequence ,Epidermal growth factor receptor ,Lung cancer ,Chromatography, High Pressure Liquid ,In Situ Hybridization, Fluorescence ,Genetics (clinical) ,Mutation ,biology ,Kinase ,Receptor Protein-Tyrosine Kinases ,Exons ,Sequence Analysis, DNA ,medicine.disease ,Protein Structure, Tertiary ,ErbB Receptors ,Protein kinase domain ,Cancer research ,biology.protein ,Molecular Medicine ,Adenocarcinoma ,Glioblastoma - Abstract
Mutations that may predict response to adenosine 5'-triphosphate (ATP)-mimetic epidermal growth factor receptor (EGFR) inhibitors occur in the EGFR kinase domain in lung adenocarcinomas and bronchioloalveolar carcinomas (BACs). Data on the frequency of EGFR mutations are sparse in other human tumors. Apart from the deletion mutant EGFRvIII, little is known about the frequency of mutations that encode for the EGFR extracellular domains II and IV that participate in receptor dimerization and formation of the tethered (autoinhibited) receptor conformation. We investigated 566 human neoplasms consisting of various histological types for mutations in exons 6, 7 (encode domain II), 14, 15 (domain IV), 18, 19, and 21 (the kinase domain) using denaturing high-performance liquid chromatography (DHPLC). Approximately 4,500 EGFR exons were screened for the presence of a mutation, and samples with an abnormal finding in DHPLC were sequenced. Only one mutation was found in the extracellular domain IV (glioblastoma), and none in domain II. Eight (11%) out of the 40 lung adenocarcinomas, or 33 BACs, investigated had exon 19 or 21 mutation in the kinase domain, but no mutations were found in other tumor types. Most of the lung cancers with mutated EGFR had three to six copies of the mutated gene in fluorescence in situ hybridization. We conclude that mutations of the EGFR kinase domain and the cysteine-rich extracellular domains are infrequent in most types of human cancer apart from lung adenocarcinoma. Mutated EGFR is usually not amplified in lung cancer.
- Published
- 2005
21. MET Receptor Tyrosine Kinase Sequence Alterations in Differentiated Thyroid Carcinoma
- Author
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Marja-Liisa Karjalainen-Lindsberg, Samuli Hemmer, Nina N. Nupponen, Heikki Joensuu, Kaarle Franssila, and Veli-Matti Wasenius
- Subjects
Male ,Pathology ,medicine.medical_specialty ,DNA Mutational Analysis ,Polymerase Chain Reaction ,Pathology and Forensic Medicine ,Immunoenzyme Techniques ,Thyroid carcinoma ,Germline mutation ,Proto-Oncogene Proteins ,Adenocarcinoma, Follicular ,Biomarkers, Tumor ,Carcinoma ,Humans ,Medicine ,Receptors, Growth Factor ,Thyroid Neoplasms ,Chromatography, High Pressure Liquid ,Germ-Line Mutation ,Aged ,DNA Primers ,Papillary renal cell carcinomas ,business.industry ,Cancer ,DNA, Neoplasm ,Middle Aged ,Proto-Oncogene Proteins c-met ,medicine.disease ,Carcinoma, Papillary ,Carcinoma, Medullary ,Adenocarcinoma ,Female ,Surgery ,Anatomy ,business ,PAX8 - Abstract
Activating mutations affecting the MET receptor tyrosine kinase are present in several types of human cancer, particularly in papillary renal cell carcinoma. Papillary thyroid carcinomas commonly express high levels of MET mRNA and protein, suggesting that increased MET signaling may be of importance in the molecular pathogenesis of differentiated thyroid carcinoma. To evaluate the role of MET mutations in thyroid carcinoma, we screened MET exons 2 to 21 for mutations in sporadic papillary, follicular, medullary, and anaplastic thyroid carcinomas using denaturing high-performance chromatography. A missense MET sequence alteration T1010I, located in exon 14 encoding for the juxtamembrane domain of MET, was found in 6 (6%) of the 104 thyroid carcinomas examined, whereas all 92 goiter samples had wild-type exon 14 (P = 0.031). Three (6%) of the 53 papillary, 2 (10%) of the 21 follicular, 1 (8%) of the 13 medullary, and none of the 17 anaplastic carcinomas studied had MET(T1010I). Four of the 6 T1010I sequence alterations were present also in the germline. MET protein expression showed no apparent association with the presence of MET(T1010I), and the clinical features of the patients with cancer with MET(T1010I) were similar to those of patients whose cancer did not harbor MET(T1010I). We conclude that MET(T1010I) sequence alteration is relatively frequent in differentiated thyroid carcinoma. The clinical and the molecular pathologic significance of this MET sequence alteration is not known.
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- 2005
22. Sequence variation in the ATP8B1 gene and intrahepatic cholestasis of pregnancy
- Author
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Kristiina Aittomäki, Jodie N. Painter, Anne Ropponen, Miia Savander, Nina N. Nupponen, Anna-Elina Lehesjoki, Seija Riikonen, and Olavi Ylikorkala
- Subjects
Male ,Heterozygote ,Genotype ,Genetic Linkage ,Physiology ,Cholestasis, Intrahepatic ,Biology ,Genetic Heterogeneity ,03 medical and health sciences ,Exon ,0302 clinical medicine ,Cholestasis ,Pregnancy ,Polymorphism (computer science) ,Genetics ,medicine ,Humans ,Genetics (clinical) ,030304 developmental biology ,Adenosine Triphosphatases ,0303 health sciences ,integumentary system ,musculoskeletal, neural, and ocular physiology ,Haplotype ,Progressive familial intrahepatic cholestasis ,Exons ,ABCB4 ,medicine.disease ,nervous system diseases ,3. Good health ,Pregnancy Complications ,Haplotypes ,Female ,030211 gastroenterology & hepatology ,sense organs ,Cholestasis of pregnancy - Abstract
Intrahepatic cholestasis of pregnancy (ICP) is a cholestatic condition that may affect women during the third trimester of pregnancy. Symptoms experienced by these women generally resolve spontaneously following delivery, but prior to delivery the fetus is at increased risk of intrauterine distress and sudden intrauterine death. The genetic etiology of most cases of ICP is unknown, although heterozygous carriers of mutations causing progressive familial intrahepatic cholestasis (PFIC) diseases may experience ICP. When examining linkage to known cholestasis genes, affected members of four Finnish ICP families shared haplotypes around ATP8B1, the gene responsible for PFIC1. This gene was subsequently screened in 176 familial and sporadic ICP patients. A total of 17 sequence changes were detected, five exonic and 12 intronic. No intronic change was associated with ICP in sporadic cases. Four intronic changes segregated with ICP in three families, a different change in each of two families and three changes in another family, although the significance of this is currently unknown. Three exonic changes were nonsynonymous, one (in exon 23) is probably a polymorphism while two predict novel amino-acid replacements (N45T and K203R). These changes, in exons 2 and 7, were detected in one individual each, and may have predisposed these individuals to ICP. In conclusion, although the exon 2 and 7 changes may have functioned as risk alleles, ATP8B1 is probably not a major gene contributing to the occurrence of ICP.
- Published
- 2005
23. Early-Onset Renal Cell Carcinoma as a Novel Extraparaganglial Component of SDHB-Associated Heritable Paraganglioma
- Author
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Maija Ht Kiuru, Charis Eng, Heikki Järvinen, Eero Pukkala, Andrzej Januszewicz, Riitta Herva, Mary Buchta, Mariola Pęczkowska, Matti Juhola, Lauri A. Aaltonen, Nina N. Nupponen, Carl Morrison, Rainer Lehtonen, Sarah R. McWhinney, Sanna K. Virta, Jukka-Pekka Mecklin, Sakari Vanharanta, and Hartmut P. H. Neumann
- Subjects
Adult ,Iron-Sulfur Proteins ,Male ,medicine.medical_specialty ,Adolescent ,SDHB ,Population ,Mutation, Missense ,Biology ,Germline ,Pheochromocytoma ,Paraganglioma ,03 medical and health sciences ,0302 clinical medicine ,Germline mutation ,Internal medicine ,Report ,medicine ,Genetics ,Humans ,Genetic Predisposition to Disease ,Genetics(clinical) ,Age of Onset ,education ,Carcinoma, Renal Cell ,Genetics (clinical) ,Germ-Line Mutation ,030304 developmental biology ,Sequence Deletion ,0303 health sciences ,education.field_of_study ,Hereditary Paraganglioma ,Base Sequence ,Siblings ,medicine.disease ,Kidney Neoplasms ,3. Good health ,Pedigree ,Succinate Dehydrogenase ,Protein Subunits ,Endocrinology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,SDHD - Abstract
Hereditary paraganglioma syndrome has recently been shown to be caused by germline heterozygous mutations in three (SDHB, SDHC, and SDHD) of the four genes that encode mitochondrial succinate dehydrogenase. Extraparaganglial component neoplasias have never been previously documented. In a population-based registry of symptomatic presentations of phaeochromocytoma/paraganglioma comprising 352 registrants, among whom 16 unrelated registrants were SDHB mutation positive, one family with germline SDHB mutation c.847-50delTCTC had two members with renal cell carcinoma (RCC), of solid histology, at ages 24 and 26 years. Both also had paraganglioma. A registry of early-onset RCCs revealed a family comprising a son with clear-cell RCC and his mother with a cardiac tumor, both with the germline SDHB R27X mutation. The cardiac tumor proved to be a paraganglioma. All RCCs showed loss of the remaining wild-type allele. Our observations suggest that germline SDHB mutations can predispose to early-onset kidney cancers in addition to paragangliomas and carry implications for medical surveillance.
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- 2004
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24. Mutational analysis of susceptibility genesRNASEL/HPC1,ELAC2/HPC2, andMSR1in sporadic prostate cancer
- Author
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Damaris Ponciano, Christiane M. Robbins, Robert L. Vessella, Mika J. Wallén, Tapio Visakorpi, John D. Carpten, Teuvo L.J. Tammela, and Nina N. Nupponen
- Subjects
Genetics ,0303 health sciences ,Cancer Research ,RNASEL Gene ,Biology ,urologic and male genital diseases ,medicine.disease ,3. Good health ,03 medical and health sciences ,Prostate cancer ,ELAC2 ,0302 clinical medicine ,Germline mutation ,medicine.anatomical_structure ,DU145 ,Prostate ,030220 oncology & carcinogenesis ,LNCaP ,medicine ,Missense mutation ,030304 developmental biology - Abstract
Three putative prostate cancer-susceptibility genes, RNASEL/HPC1 at 1q24, MSR1 at 8p22, and ELAC2/HPC2 at 17p11, have recently been identified. Our objective was to investigate somatic mutations in these genes in sporadic prostate cancer. We analyzed 39 clinical prostate cancer specimens, 10 prostate cancer xenografts (LuCaP series), and 4 prostate cancer cell lines (LNCaP, DU145, PC-3, and MPC-3) for genetic changes using denaturing high-performance liquid chromatography and direct sequencing in order to screen the whole coding regions of RNASEL and MSR1, as well as exons 7 and 17 of ELAC2. The known 471delAAAG truncating mutation was found in the RNASEL gene in cell line LNCaP. The only new missense variation in RNASEL, Gly296Val, was found in cell line DU145, but not in any other samples. RNASEL and ELAC2 also showed the common missense polymorphic changes. A previously reported truncating mutation (Arg293X) was found in MSR1 in the germ line of one individual. Our results indicate that inactivation of the RNASEL, ELAC2, or MSR1 genes by somatic mutation is a rare phenomenon in sporadic prostate cancer.
- Published
- 2003
25. Human pHyde is not a classical tumor suppressor gene in prostate cancer
- Author
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Teuvo L.J. Tammela, Kati P. Porkka, Tapio Visakorpi, Robert L. Vessella, and Nina N. Nupponen
- Subjects
Male ,PCA3 ,Cancer Research ,Pathology ,medicine.medical_specialty ,Neoplasms, Hormone-Dependent ,Tumor suppressor gene ,DNA Mutational Analysis ,Transplantation, Heterologous ,Prostatic Hyperplasia ,Cell Cycle Proteins ,Biology ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,DU145 ,Prostate ,LNCaP ,Gene expression ,Tumor Cells, Cultured ,medicine ,Animals ,Humans ,Genes, Tumor Suppressor ,RNA, Messenger ,In Situ Hybridization, Fluorescence ,DNA Primers ,030304 developmental biology ,Oncogene Proteins ,0303 health sciences ,medicine.diagnostic_test ,Reverse Transcriptase Polymerase Chain Reaction ,Prostatic Neoplasms ,DNA ,Blotting, Northern ,medicine.disease ,Rats ,3. Good health ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Oxidoreductases ,Fluorescence in situ hybridization - Abstract
A novel putative tumor suppressor gene, pHyde, was recently cloned from rat prostate. The rat gene has been shown to inhibit prostate cancer cell proliferation both in vitro and in vivo. However, the role of human pHyde in prostate cancer has not been studied before. Here, we analyzed human prostate cancer cell lines (LNCaP, DU145, PC-3, 22Rv1), xenografts (LuCaP 23.1, 35, 41, 49, 58, 69, 70 and 73) and clinical prostate carcinomas for genetic alterations and expression of pHyde. The expression of pHyde in normal human tissues as well as in prostate cancer was studied by Northern analysis and real-time quantitative RT-PCR. It was ubiquitously expressed in all normal tissues analyzed. Although, the expression was significantly (p=0.007) lower in poorly differentiated than in well and moderately differentiated carcinomas, there were no differences in the expression levels between benign prostate hyperplasia, untreated primary and recurrent hormone-refractory prostate carcinomas (p=0.607). Altogether, missense mutations were detected in 2 out of 68 samples studied (∼3%) by denaturing high-performance liquid chromatography (DHPLC) and sequencing. One of the samples with the mutation also exhibited a loss of a gene copy by fluorescence in situ hybridization (FISH). This was the only sample that exhibited a genetic alteration in both alleles, suggesting that the human pHyde is not a classical prostate tumor suppressor gene. The reduced expression of the gene found in some tumors warrant further studies. © 2003 Wiley-Liss, Inc.
- Published
- 2003
26. Little evidence for involvement ofMLH3in colorectal cancer predisposition
- Author
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Reijo Salovaara, Heikki Järvinen, Pertti Sistonen, Virpi Launonen, Jukka-Pekka Mecklin, Auli Karhu, Nina N. Nupponen, Lauri A. Aaltonen, Rainer Lehtonen, Päivi Peltomäki, Tuija Hienonen, and Päivi Laiho
- Subjects
Genetics ,congenital, hereditary, and neonatal diseases and abnormalities ,0303 health sciences ,Cancer Research ,MLH3 ,Biology ,MLH1 ,digestive system diseases ,3. Good health ,Frameshift mutation ,MSH6 ,03 medical and health sciences ,0302 clinical medicine ,Germline mutation ,Oncology ,MSH2 ,030220 oncology & carcinogenesis ,PMS2 ,Missense mutation ,neoplasms ,030304 developmental biology - Abstract
Mutations in the DNA MMR genes MSH2, MLH1, MSH6 and PMS2 underlie a large subset of HNPCC cases, and a hallmark of the tumors is MSI. In many HNPCC families, however, a causative mutation has not been found. Therefore, the involvement of additional, thus far unknown, genes in MSI as well as MSS colorectal tumor predisposition is possible. The role of a relatively recently cloned MMR gene, MLH3, in familial CRC has been studied; but the results appear somewhat conflicting. To further evaluate the role of MLH3 in CRC predisposition, we analyzed 30 Finnish CRC cases for germline mutations by sequencing. These cases were selected from a large series of Finnish CRC patients, to match features previously proposed to associate with MLH3 germline defects. We found 5 missense variants, 4 of which were also found in Finnish cancer-free controls. The only remaining variant does not appear to be an attractive candidate for a disease-associated mutation because the amino acid change is located outside the conserved residues. We also screened for the previously reported variants, including a frameshift change, the most likely pathogenic MLH3 mutation observed so far. The frameshift was not present in the 30 CRC cases or in 700 cancer-free controls. While it is a difficult task to exclude a role of MLH3 in HNPCC, our study could not confirm a role for MLH3 in CRC predisposition. © 2003 Wiley-Liss, Inc.
- Published
- 2003
27. Germline Alterations of the RNASEL Gene, a Candidate HPC1 Gene at 1q25, in Patients and Families with Prostate Cancer
- Author
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Tarja Ikonen, Annika Rökman, Olli Kallioniemi, Nina Mononen, Pasi A. Koivisto, Teuvo L.J. Tammela, Eija H. Seppälä, Mika P. Matikainen, John D. Carpten, Jeffrey M. Trent, Nina N. Nupponen, Ville Autio, Joan E. Bailey-Wilson, and Johanna Schleutker
- Subjects
Male ,DNA Mutational Analysis ,Mutation, Missense ,Biology ,urologic and male genital diseases ,MSR1 ,Prostate cancer ,Germline mutation ,Gene Frequency ,Endoribonucleases ,Genetics ,medicine ,Humans ,Missense mutation ,Genetics(clinical) ,Age of Onset ,Mutation frequency ,Allele frequency ,Finland ,Germ-Line Mutation ,Polymorphism, Single-Stranded Conformational ,Genetics (clinical) ,Prostatic Neoplasms ,RNASEL Gene ,Articles ,medicine.disease ,Pedigree ,Chromosomes, Human, Pair 1 ,Female ,Age of onset - Abstract
The RNASEL gene (2',5'-oligoisoadenylate-synthetase dependent) encodes a ribonuclease that mediates the antiviral and apoptotic activities of interferons. The RNASEL gene maps to the hereditary-prostate-cancer (HPC)-predisposition locus at 1q24-q25 (HPC1) and was recently shown to harbor truncating mutations in two families with linkage to HPC1. Here, we screened for RNASEL germline mutations in 66 Finnish patients with HPC, and we determined the frequency of the changes in the index patients from 116 families with HPC, in 492 patients with unselected prostate cancer (PRCA), in 223 patients with benign prostatic hyperplasia (BPH), and in 566 controls. A truncating mutation, E265X, was found in 5 (4.3%) of the 116 patients from families with HPC. This was significantly higher (odds ratio [OR] =4.56; P=.04) than the frequency of E265X in controls (1.8%). The highest mutation frequency (9.5%) was found in patients from families with four or more affected members. Possible segregation was detected only in a single family. However, the median age at disease onset for E265X carriers was 11 years less than that for noncarriers in the same families. In addition, of the four missense variants found, R462Q showed an association with HPC (OR=1.96; P=.07). None of the variants showed any differences between controls and either patients with BPH or patients with PRCA. We conclude that, although RNASEL mutations do not explain disease segregation in Finnish families with HPC, the variants are enriched in families with HPC that include more than two affected members and may also be associated with the age at disease onset. This suggests a possible modifying role in cancer predisposition. The impact that the RNASEL sequence variants have on PRCA burden at the population level seems small but deserves further study.
- Published
- 2002
28. Amplification of EIF3S3 Gene Is Associated with Advanced Stage in Prostate Cancer
- Author
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Outi R. Saramäki, Nina N. Nupponen, Ola Bratt, Lukas Bubendorf, Niels Willi, Tapio Visakorpi, Thomas C. Gasser, and Pasi A. Koivisto
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Eukaryotic Initiation Factor-3 ,Gene Dosage ,Biology ,Gene dosage ,Pathology and Forensic Medicine ,Prostate cancer ,Peptide Initiation Factors ,Prostate ,medicine ,Humans ,In Situ Hybridization, Fluorescence ,Aged ,Neoplasm Staging ,Aged, 80 and over ,Tissue microarray ,medicine.diagnostic_test ,Gene Amplification ,Prostatic Neoplasms ,Regular Article ,Middle Aged ,medicine.disease ,Survival Analysis ,Survival Rate ,medicine.anatomical_structure ,Cancer research ,Adenocarcinoma ,Oncogene MYC ,Comparative genomic hybridization ,Fluorescence in situ hybridization - Abstract
Gain of the long arm of chromosome 8 (8q) is one of the most common gains found in the advanced prostate cancer by comparative genomic hybridization. We have previously identified a putative target gene for the 8q gain, EIF3S3, that encodes a p40 subunit of eukaryotic translation initiation factor 3 (eIF3). Here, we studied the frequency of the EIF3S3 amplification in different stages of prostate cancer and co-amplification of EIF3S3 and oncogene MYC. In addition, prognostic utility of the EIF3S3 copy number alteration was evaluated. The analyses were done with fluorescence in situ hybridization and tissue microarray technology. High-level amplification of EIF3S3 was found in 11 of 125 (9%) of pT1/pT2 tumors, 12 of 44 (27%) of pT3/pT4 tumors, and 8 of 37 (22%) of lymph node metastases as well as in 26 of 78 (33%) and 15 of 30 (50%) of hormone refractory locally recurrent tumors and metastases, respectively. The amplification was associated with high Gleason score (P < 0.001). One of the 79 tumors with EIF3S3 amplification had only two copies of MYC, whereas all tumors with amplification of MYC had also amplification of EIF3S3 indicating common co-amplification of the genes. Gain of EIF3S3 was associated with poor cancer-specific survival in incidentally found prostate carcinomas (P = 0.023). In the analyses of prostatectomy-treated patients, the amplification was not statistically significantly associated with progression-free time. In conclusion, amplification of EIF3S3 gene is common in late-stage prostate cancer suggesting that it may be functionally involved in the progression of the disease.
- Published
- 2001
29. Amplification of hypoxia-inducible factor 1α gene in prostate cancer
- Author
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Outi R. Saramäki, Ola Bratt, Nina N. Nupponen, Kimmo Savinainen, and Tapio Visakorpi
- Subjects
Male ,PCA3 ,Cancer Research ,HIF1A Gene ,Gene Dosage ,Biology ,Gene dosage ,Prostate cancer ,Gene duplication ,LNCaP ,Tumor Cells, Cultured ,Genetics ,medicine ,Humans ,Molecular Biology ,In Situ Hybridization, Fluorescence ,Chromosomes, Human, Pair 14 ,Helix-Loop-Helix Motifs ,Gene Amplification ,Chromosome Mapping ,Nuclear Proteins ,Nucleic Acid Hybridization ,Prostatic Neoplasms ,Chromoplexy ,Hypoxia-Inducible Factor 1, alpha Subunit ,medicine.disease ,DNA-Binding Proteins ,HIF1A ,Cancer research ,Hypoxia-Inducible Factor 1 ,Transcription Factors - Abstract
Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that regulates the expression of genes associated with adaptation to reduced oxygen pressure. Increased expression of HIF-1alpha gene (HIF1A) has been found in the majority of prostate carcinomas. In addition, the PC-3 prostate cancer cell line has been shown to express the gene even under normoxic conditions. By comparative genomic hybridization (CGH), we have earlier shown that the PC-3 cell line contains a high-level amplification in the chromosomal region harboring the HIF1A gene. Here, we first fine mapped the gene to locus 14q23 by fluorescence in situ hybridization (FISH). The gene was then shown to be highly amplified in the PC-3 cell line. Subsequently, the copy number of the HIF1A gene was studied in 5 other prostate cancer cell lines (LNCaP, DU-145, NCI-H660, Tsu-Pr, JCA-1) and in 117 prostate tumors representing both hormone-dependent and -refractory disease as well as primary and metastatic lesions. No high-level amplifications of the HIF1A gene were found. Additional copies of the gene were seen in all of the cell lines and in 36% of the tumors. There was no association between the tumor type and the copy number alterations of the gene. In conclusion, high-level amplification of the HIF1A gene may explain the overexpression of the gene in the PC-3 prostate cancer cell line. However, such high-level amplification seems to be very rare in prostate cancer.
- Published
- 2001
30. 5q11, 8p11, and 10q22 are recurrent chromosomal breakpoints in prostate cancer cell lines
- Author
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Yi Pan, Catharina Larsson, Jorma Isola, Weng-Onn Lui, Nina N. Nupponen, Tapio Visakorpi, Soili Kytölä, and Ulf S.R. Bergerheim
- Subjects
Genetic Markers ,Male ,Cancer Research ,DNA Mutational Analysis ,Chromosomal translocation ,Biology ,Translocation, Genetic ,Prostate cancer ,DU145 ,LNCaP ,Tumor Cells, Cultured ,Genetics ,medicine ,Humans ,Chromosomes, Human, Pair 10 ,Breakpoint ,Nucleic Acid Hybridization ,Prostatic Neoplasms ,Chromosome Breakage ,Karyotype ,Chromoplexy ,Genes, p53 ,medicine.disease ,Molecular biology ,Karyotyping ,Chromosomes, Human, Pair 5 ,Chromosome Deletion ,Chromosomes, Human, Pair 8 ,Comparative genomic hybridization - Abstract
Prostate cancer cell lines have been widely used as model systems characterizing pathogenetic, functional, and therapeutic aspects of prostate cancer development. However, their chromosomal compositions are poorly characterized. In this study, five prostate cancer cell lines-TSU-Pr1, JCA-1, NCI-H660, ALVA-31, and PPC-1-were investigated by G-banding, comparative genomic hybridization (CGH), and spectral karyotyping. The results were combined with our previous findings in the prostate cancer cell lines PC-3, DU145, and LNCaP. By comparative genomic hybridization (CGH), the most frequent losses were observed at 13q, 8p, 9p, and 4q, whereas gains were most commonly seen at 8q, 10q, and 18p. The composite karyotypes were characterized by multiple numerical and structural chromosomal aberrations. Recurrent breakpoints at 5q11, 8p11, and 10q22 were observed to participate in deletion and translocation events in five of the cell lines, suggesting the importance of tumor suppressor and/or oncogenes in these regions. ALVA-31 and PPC-1 shared nine identical derivative chromosomes, two of which have also been detected in PC-3. In addition, the identification of the same homozygous deletion at D10S541 and of an identical TP53 gene mutation in all three cell lines suggests a common origin of these cell lines.
- Published
- 2001
31. Identification of Novel Transcription Factor-like Gene from Human Intestinal Cells
- Author
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Mauno Vihinen, Nina N. Nupponen, Markku Mäki, Tuula Halttunen, Katri Lindfors, Tapio Visakorpi, Paula Huotari, and Heikki Kainulainen
- Subjects
Molecular Sequence Data ,Biophysics ,Biology ,Polymerase Chain Reaction ,Biochemistry ,Cell Line ,Transforming Growth Factor beta ,Complementary DNA ,TGF beta signaling pathway ,Humans ,Tissue Distribution ,Amino Acid Sequence ,Molecular Biology ,Transcription factor ,Gene ,Regulation of gene expression ,Binding Sites ,Base Sequence ,Sequence Homology, Amino Acid ,Chromosome Mapping ,Cell Biology ,Transforming growth factor beta ,Molecular biology ,Intestines ,Repressor Proteins ,Gene Expression Regulation ,Cell culture ,Transforming growth factor, beta 3 ,biology.protein ,Apoptosis Regulatory Proteins ,Chromosomes, Human, Pair 17 ,Transcription Factors - Abstract
Intestinal crypt epithelial T84 cells form luminal structures and differentiate to intestinal enterocyte-like cells in response to IMR-90 fibroblast-secreted transforming growth factor-beta when grown within three-dimensional collagen gel. In search of TGF-beta regulated genes involved in this differentiation process, we isolated a TGF-beta downregulated cDNA, human homologue of rat apoptosis antagonising transcription factor that codes for a 560-amino-acid protein. Human AATF-mRNA was expressed at high levels in human brain, heart, thymus, kidney, and placenta while in skeletal muscle and colon the expression was lower. The gene was mapped to chromosome 17q11.2-q12.
- Published
- 2000
32. Mapping the amplification ofEIF3S3 in breast and prostate cancer
- Author
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Jorma Isola, Tapio Visakorpi, and Nina N. Nupponen
- Subjects
Cancer Research ,Expressed sequence tag ,medicine.diagnostic_test ,In situ hybridization ,Biology ,medicine.disease ,Molecular biology ,Gene expression profiling ,Prostate cancer ,Genetic marker ,Genetics ,medicine ,Gene ,Comparative genomic hybridization ,Fluorescence in situ hybridization - Abstract
Gain of chromosome arm 8q is a frequent genetic alteration in breast and prostate cancer. Two amplified subregions, 8q21 and 8q23-24, have been identified with comparative genomic hybridization (CGH). We have recently demonstrated that the EIF3S3 (eIF3-p40) gene, located at 8q23, is often amplified and overexpressed in both breast and prostate cancer. Here, we used fluorescence in situ hybridization (FISH) to map the amplified region around EIF3S3 in primary breast cancers and cell lines. The size of the common highly amplified region was about 2.5 Mb between the markers D8S1668 and WI-7959. Next, we analyzed the expression of all expressed sequence tags (ESTs) located within and near this region by RNA slot blot hybridization. In addition to EIF3S3, three anonymous ESTs and EXT1 were found to be highly expressed in cancer cell lines with the amplification at 8q23-q24. However, the anonymous ESTs were located outside the minimal highly amplified region and EXT1 was overexpressed only in one of the cancer cell lines with 8q amplification. Since EIF3S3 was the only consistently overexpressed gene located in the minimal highly amplified region, it is the strongest candidate target gene for 8q23-q24 amplification.
- Published
- 2000
33. Molecular cytogenetics of prostate cancer
- Author
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Nina N. Nupponen and Tapio Visakorpi
- Subjects
Genetics ,medicine.medical_specialty ,Histology ,medicine.diagnostic_test ,Cytogenetics ,Biology ,medicine.disease ,Malignancy ,Molecular cytogenetics ,Loss of heterozygosity ,Medical Laboratory Technology ,Prostate cancer ,medicine ,Anatomy ,Instrumentation ,Gene ,Comparative genomic hybridization ,Fluorescence in situ hybridization - Abstract
Prostate cancer is the most common malignancy among men in Western industrialized countries. The molecular pathogenesis of the disease is poorly known. Over the past 10 years, chromosomal aberrations in prostate cancer have been studied with several techniques, such as loss of heterozygosity (LOH), classical cytogenetics, and molecular cytogenetics, namely with fluorescence in situ hybridization (FISH) and comparative genomic hybridization (CGH). These analyses, especially those performed by CGH, have enabled the distinction of the predominant chromosomal regions of involvement in prostate cancer. Studies have shown that the most common chromosomal alterations in prostate cancer are losses at 1p, 6q, 8p, 10q, 13q, 16q, and 18q and gains at 1q, 2p, 7, 8q, 18q, and Xq. Fluorescence in situ hybridization (FISH) has been used to identify the target genes for some of these chromosomal alterations. For example, amplifications of AR (at Xq12), MYC (8q24), and EIF3S3 (8q23) have been found in a large fraction of hormone-refractory prostate cancer by FISH. However, many of the critical oncogenes and tumor suppressor genes located in the altered chromosomal regions have not yet been identified.
- Published
- 2000
34. Multiple copies of mutantBRCA1 andBRCA2 alleles in breast tumors from germ-line mutation carriers
- Author
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Nina N. Nupponen, Synnöve Staff, Jorma Isola, Minna Tanner, and Åke Borg
- Subjects
Genetics ,Cancer Research ,endocrine system diseases ,Wild type ,Locus (genetics) ,Biology ,Null allele ,Molecular biology ,Germline mutation ,Allelic Imbalance ,Gene duplication ,Gene conversion ,Allele ,skin and connective tissue diseases - Abstract
Inactivation of the BRCA1 and BRCA2 breast cancer susceptibility genes has been reported to occur via a germ-line mutation of one allele and a somatic loss of the remaining wild-type allele. We investigated the genetic mechanisms behind the second event in breast tumors from 17 BRCA1 and eight BRCA2 germ-line mutation carriers, as compared with 21 sporadic breast tumors. Microsatellite markers intragenic or in close proximity to both genes were used to analyze imbalances between the mutant and wild-type alleles. The actual and relative gene copy numbers were scored by fluorescence in situ hybridization (FISH) analysis of tumor cells using locus and centromere specific probes. All but one of the informative BRCA1 and BRCA2 tumors exhibited allelic imbalance and loss of the corresponding wild type allele. In contrast to sporadic tumors, however, where allelic imbalance at the BRCA1 and BRCA2 loci correlated well with relative copy number losses by FISH, a simple reduction to a single copy (average copy number ratio 2:1) was found in only two BRCA1 (12%) and four BRCA2 (50%) tumors. The majority of BRCA1 and BRCA2 tumors showed a copy number reduction (relative to reference probe with ratios 4:2, 3:2, 4:3) at corresponding loci, suggesting that a specific physical deletion of the wild-type BRCA gene allele has been followed by a duplication of the remaining mutant allele via polyploidization. Several tumors contained multiple copies of BRCA1 and BRCA2 genes without relative copy number changes, implying that loss of wild-type alleles is executed by alternative mechanisms such as mitotic recombination, non-disjunctional chromosomal loss with or without reduplication, or by gene conversion. A paradoxical relative copy number gain of the mutant allele was evident in three BRCA1 tumors (18%), which could be of biological relevance if a dominant negative or gain-of-function model was ascribed for certain BRCA1 mutants. Our results indicate that complex genetic alterations are operational at the BRCA1 and BRCA2 loci in tumors from genetically predisposed individuals.
- Published
- 2000
35. Cyclin D1 expression in astrocytomas is associated with cell proliferation activity and patient prognosis
- Author
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Heikki Helin, Pauli Helén, Juha Kononen, Kirsi Syrjäkoski, Nina N. Nupponen, Satu-Leena Sallinen, Immo Rantala, Pauli Sallinen, and Hannu Haapasalo
- Subjects
Pathology ,medicine.medical_specialty ,medicine.diagnostic_test ,Cell growth ,Astrocytoma ,In situ hybridization ,Biology ,Cell cycle ,medicine.disease ,Pathology and Forensic Medicine ,Cyclin D1 ,medicine ,Cancer research ,Immunohistochemistry ,Mitosis ,Fluorescence in situ hybridization - Abstract
An important positive regulator of the cell cycle, cyclin D1, is often amplified and overexpressed in malignancies. Cyclin D1 aberrations were analysed in grade II–IV astrocytomas by fluorescence in situ hybridization (FISH), mRNA in situ hybridization and immunohistochemistry. Proliferation activity was determined by Ki-67MIB-1 immunolabelling and mitotic counting. High cyclin D1 expression was observed in grade IV astrocytomas (grades II–III versus grade IV; mRNA expression: p
- Published
- 1999
36. Vol. 35, 1999
- Author
-
Colin W. Bayne, Yves Fradet, Polly Feigl, A.R. Zlotta, Peter Boyle, Roxann M. Neumann, Junqi Qian, Ronald A. Lubet, Levy Kopelovitch, Thomas Putz, C.C. Schulman, B. Têtu, Donna M. Peehl, Gary J. Kelloff, Don W. W. Newling, Margaret Ross, Ferran Algaba, Peter Whelan, Judd W. Moul, D. Altshuler, B.E. Henderson, M. Marshall, James A. Crowell, David L. McCormick, Jagat Ghosh, C.L. Pearce, Gianluca Severi, E. Lander, R.K. Ross, Vinicius Duval da Silva, T.H. Van Der Kwast, G. Daley, K. Griffiths, Andrew C.B. Cato, Rodolfo Montironi, Nina N. Nupponen, Alfred Hobisch, Ramak Shadmani, M. Markiewicz, Peter W. Hamilton, Georg Bartsch, E.D. Crawford, Wim J. Kirkels, Lynne Moore, Iris E. Eder, Charles W. Boone, L. Denis, Liang Cheng, J.K.V. Reichardt, Ian M. Thompson, Helmut Klocker, P. Bretsky, Marina Scarpelli, Charles E. Myers, David J. Waters, Marion J.G. Bussemakers, R. Lieberman, Robert B. Jenkins, Caroline C. Sigman, K.V.N. Rao, Isabelle Bairati, Vernon E. Steele, Zoran Culig, M. Studen, U. Manne, Fritz H. Schröder, D. Perkins, G.A. Coetzee, Peter H. Bartels, David G. Bostwick, A. Reissigl, Wael Sakr, W. Grizzle, D. Urban, Peter Ekman, François Meyer, Claudia Nessler-Menardi, L.N. Kolonel, Hubert G. Bartels, Deborah Thompson, M.S. Morton, H. Volgger, Michael B. Sporn, W. Horninger, C.A. Coltman, G. Bartsch, Heike Peterziel, R. Myers, H. Klocker, Adrian P.M. van der Meijden, J. Mohler, F. Labrie, Ronald Lieberman, H. Rogatsch, John Rietbergen, Fouad K. Habib, G. Kelloff, Richard Sylvester, Laurence Collette, Maarten C. Bosland, Winfred A. Malone, H. Weiss, Ries Kranse, Robert F. Hoedemaeker, Tapio Visakorpi, and Linda Vaught
- Subjects
business.industry ,Urology ,Library science ,Medicine ,business - Published
- 1999
37. Prostatitis and Urethritis
- Author
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Don W. W. Newling, R.K. Ross, John Rietbergen, Ronald A. Lubet, T.H. Van Der Kwast, Fouad K. Habib, Charles W. Boone, Junqi Qian, David L. McCormick, M.S. Morton, Vinicius Duval da Silva, Isabelle Bairati, Levy Kopelovitch, Thomas Putz, Robert F. Hoedemaeker, H. Klocker, Adrian P.M. van der Meijden, J. Mohler, J.K.V. Reichardt, Tapio Visakorpi, Peter Whelan, L. Denis, M. Studen, Lynne Moore, Rodolfo Montironi, C.C. Schulman, Caroline C. Sigman, M. Markiewicz, Peter W. Hamilton, Vernon E. Steele, D. Perkins, F. Labrie, G. Bartsch, Marina Scarpelli, E.D. Crawford, Ronald Lieberman, H. Rogatsch, Alfred Hobisch, G. Kelloff, B. Têtu, Heike Peterziel, L.N. Kolonel, Colin W. Bayne, W. Grizzle, R. Myers, James A. Crowell, Nina N. Nupponen, G.A. Coetzee, Richard Sylvester, Margaret Ross, Laurence Collette, Yves Fradet, Linda Vaught, A.R. Zlotta, Peter Boyle, B.E. Henderson, David G. Bostwick, Winfred A. Malone, H. Weiss, R. Lieberman, Ries Kranse, Judd W. Moul, Ramak Shadmani, K.V.N. Rao, P. Bretsky, Liang Cheng, Polly Feigl, Zoran Culig, Andrew C.B. Cato, G. Daley, Georg Bartsch, Robert B. Jenkins, Ferran Algaba, Charles E. Myers, Roxann M. Neumann, Jagat Ghosh, A. Reissigl, Wael Sakr, Gianluca Severi, Michael B. Sporn, Claudia Nessler-Menardi, U. Manne, D. Urban, Peter Ekman, Maarten C. Bosland, Donna M. Peehl, Gary J. Kelloff, François Meyer, C.A. Coltman, Iris E. Eder, W. Horninger, Deborah Thompson, H. Volgger, Hubert G. Bartels, D. Altshuler, Fritz H. Schröder, Wim J. Kirkels, Ian M. Thompson, David J. Waters, Marion J.G. Bussemakers, C.L. Pearce, E. Lander, M. Marshall, Peter H. Bartels, Helmut Klocker, and K. Griffiths
- Subjects
medicine.medical_specialty ,business.industry ,Urology ,Medicine ,Prostatitis ,Urethritis ,business ,medicine.disease - Published
- 1999
38. Genetic Aberrations in Hypodiploid Breast Cancer
- Author
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Jorma Isola, Minna Tanner, Tanja Pejovic, Ritva Karhu, Nina N. Nupponen, Mårten Fernö, Åke Borg, Dick Killander, and Bo Baldetorp
- Subjects
medicine.diagnostic_test ,Biology ,medicine.disease ,Molecular biology ,Pathology and Forensic Medicine ,Cyclin D1 ,Breast cancer ,Gene duplication ,medicine ,Hypodiploidy ,Hyperdiploidy ,Ploidy ,Comparative genomic hybridization ,Fluorescence in situ hybridization - Abstract
The evolution of somatic genetic aberrations in breast cancer has remained poorly understood. The most common chromosomal abnormality is hyperdiploidy, which is thought to arise via a transient hypodiploid state. However, hypodiploidy persists in 1 to 2% of breast tumors, which are characterized by a poor prognosis. We studied the genetic aberrations in 15 flow cytometrically hypodiploid breast cancers by comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH). Surprisingly, numerous copy number gains were detected in addition to the copy number losses. The number of gains per tumor was 4.3 +/- 3.2 and that of losses was 4.5 +/- 3.3 (mean +/- SD), which is similar to that previously observed in hyperdiploid breast cancers. Gains at chromosomes or chromosomal regions at 11q13, 1q, 19, and 16p and losses of 2q, 4, 6q, 9p, 13, and 18 were most commonly observed. Compared with unselected breast carcinomas, hypodiploid tumors showed certain differences. Loss of chromosome 4 (53%) and gain of 11q13 (60%) were significantly more common in hypodiploid tumors. The gain at 11q13 was found by FISH to harbor amplification of the Cyclin D1 oncogene, which is therefore three to four times more common in hypodiploid than in unselected breast cancers (15 to 20%). Structural chromosomal aberrations (such as Cyclin D1 amplification) were present both in diploid and hypodiploid tumor cell populations, as assessed by FISH and CGH after flow cytometric sorting. Together these results indicate that hypodiploid tumors form a distinct genetic entity of invasive breast cancer, although they probably share a common genetic evolution pathway where structural chromosomal aberrations precede gross DNA ploidy changes.
- Published
- 1998
39. Prognostic but not predictive role of platelet-derived growth factor receptors in patients with recurrent glioblastoma
- Author
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Heikki Joensuu, Andreas von Deimling, Malin Jarvius, Monica Nistér, Arne Östman, Gregor Dresemann, Janna Paulsson, Ola Söderberg, Werner Paulus, Marjut Puputti, Martin Hasselblatt, Nina N. Nupponen, and Maja Bradic Lindh
- Subjects
Male ,Cancer Research ,Platelet-derived growth factor ,IDH1 ,Receptor, Platelet-Derived Growth Factor alpha ,Combination therapy ,medicine.drug_class ,Tyrosine-kinase inhibitor ,Piperazines ,Hydroxycarbamide ,Immunoenzyme Techniques ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Growth factor receptor ,hemic and lymphatic diseases ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Hydroxyurea ,Phosphorylation ,neoplasms ,030304 developmental biology ,0303 health sciences ,biology ,Brain Neoplasms ,Imatinib ,Middle Aged ,Prognosis ,3. Good health ,Survival Rate ,Pyrimidines ,Oncology ,chemistry ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Benzamides ,biology.protein ,Cancer research ,Imatinib Mesylate ,Female ,Neoplasm Recurrence, Local ,Glioblastoma ,Platelet-derived growth factor receptor ,medicine.drug - Abstract
Platelet-derived growth factor receptor (PDGFR) signaling has been implicated in the pathogenesis of glioblastomas and represents a target for the tyrosine kinase inhibitor imatinib. To examine the prognostic or predictive role of PDGFRs in recurrent glioblastomas, expression was examined in tumor samples of 101 patients of CSTI571BDE40, a randomized trial comparing hydroxyurea monotherapy and a combination of hydroxyurea and imatinib. Furthermore, PDGFRα phosphorylation was investigated using in situ proximity ligation assay. PDGFRα protein was expressed in 33% of tumors and was associated with male sex, young age, presence of R132H mutated isocitrate dehydrogenase 1 protein and short median survival (142 vs. 187 days, p = 0.028). Tumor PDGFRα phosphorylation was also associated with short survival (p = 0.030). The subset of patients with PDGFRα positive glioblastoma did not have longer survival on treatment with hydroxyurea and imatinib compared with hydroxyurea monotherapy. In conclusion, both PDGFRα protein expression and phosphorylation status had a prognostic role in recurrent glioblastomas but did not define a group that showed benefit from the combination therapy consisting of hydroxyurea and imatinib.
- Published
- 2010
40. The tyrosine kinase c-Abl promotes proliferation and is expressed in atypical teratoid and malignant rhabdoid tumors
- Author
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Annariikka Roselli, Nina N. Nupponen, Ivo Leuschner, Björn Koos, Henning Lünenbürger, Sonja Mertsch, Astrid Jeibmann, Martin Hasselblatt, Werner Paulus, and Michael C. Frühwald
- Subjects
Cancer Research ,medicine.drug_class ,Tyrosine-kinase inhibitor ,Piperazines ,03 medical and health sciences ,0302 clinical medicine ,Growth factor receptor ,Cell Line, Tumor ,Medicine ,Humans ,Proto-Oncogene Proteins c-abl ,Protein Kinase Inhibitors ,Rhabdoid Tumor ,030304 developmental biology ,Cell Proliferation ,0303 health sciences ,ABL ,business.industry ,Cancer ,Imatinib ,Protein-Tyrosine Kinases ,medicine.disease ,Kidney Neoplasms ,3. Good health ,Teratoid tumor ,Imatinib mesylate ,Pyrimidines ,Oncology ,030220 oncology & carcinogenesis ,Benzamides ,Cancer research ,Imatinib Mesylate ,business ,Tyrosine kinase ,medicine.drug - Abstract
BACKGROUND: Atypical teratoid/rhaboid tumors (AT/RTs) and extracranial malignant rhabdoid tumors are highly malignant neoplasms with a dismal prognosis. These tumors predominantly affect infants and targeted, adjuvant treatment approaches would be highly desirable. METHODS: In the current study, the authors investigated the expression and functional role of tyrosine kinases in 2 malignant rhabdoid tumor cell lines (A204 and G401) and in a series of 5 AT/RTs and 18 malignant rhabdoid tumors (13 rhabdoid tumors of the kidney and 5 extrarenal rhabdoid tumors). RESULTS: Both cell lines consistently expressed the tyrosine kinase c-Abl, which promoted proliferation as assessed by small interfering RNA knockdown. Blockage of c-Abl using the tyrosine kinase inhibitor imatinib resulted in reduced cellular growth in both cell lines. Furthermore, c-Abl was expressed in all rhabdoid tumors, whereas expression of platelet-derived growth factor receptor subtypes alpha and beta was infrequent and c-Kit expression was absent. CONCLUSIONS: The current data pointed toward a role for c-Abl in the biology of malignant rhabdoid tumors and provided a rationale for the investigation of tyrosine kinase inhibitors that target c-Abl for the treatment of these aggressive tumors. Cancer 2010. © 2010 American Cancer Society
- Published
- 2010
41. Subject Index Vol. 35, 1999
- Author
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Colin W. Bayne, G. Bartsch, Linda Vaught, Robert B. Jenkins, Roxann M. Neumann, A. Reissigl, Wael Sakr, Rodolfo Montironi, Junqi Qian, Heike Peterziel, L. Denis, M.S. Morton, Levy Kopelovitch, Lynne Moore, Thomas Putz, Hubert G. Bartels, D. Urban, Peter Ekman, M. Markiewicz, Peter W. Hamilton, R. Myers, Gianluca Severi, Donna M. Peehl, Gary J. Kelloff, Peter Whelan, M. Marshall, Charles W. Boone, Georg Bartsch, M. Studen, K. Griffiths, Iris E. Eder, A.R. Zlotta, Charles E. Myers, Peter Boyle, Deborah Thompson, Alfred Hobisch, Helmut Klocker, E.D. Crawford, W. Horninger, R. Lieberman, Caroline C. Sigman, David L. McCormick, Ronald A. Lubet, Winfred A. Malone, David J. Waters, Marion J.G. Bussemakers, D. Altshuler, G. Daley, H. Weiss, Polly Feigl, Marina Scarpelli, K.V.N. Rao, Vernon E. Steele, Ries Kranse, H. Volgger, John Rietbergen, P. Bretsky, Claudia Nessler-Menardi, Ferran Algaba, Isabelle Bairati, B.E. Henderson, Fouad K. Habib, G.A. Coetzee, Jagat Ghosh, D. Perkins, H. Klocker, Peter H. Bartels, David G. Bostwick, Adrian P.M. van der Meijden, J. Mohler, Zoran Culig, Robert F. Hoedemaeker, W. Grizzle, Tapio Visakorpi, Nina N. Nupponen, F. Labrie, Ronald Lieberman, Laurence Collette, G. Kelloff, U. Manne, H. Rogatsch, R.K. Ross, François Meyer, L.N. Kolonel, Richard Sylvester, C.L. Pearce, E. Lander, Michael B. Sporn, Fritz H. Schröder, C.A. Coltman, J.K.V. Reichardt, Wim J. Kirkels, C.C. Schulman, Maarten C. Bosland, B. Têtu, Ian M. Thompson, Yves Fradet, James A. Crowell, Don W. W. Newling, T.H. Van Der Kwast, Judd W. Moul, Ramak Shadmani, Andrew C.B. Cato, Vinicius Duval da Silva, Margaret Ross, and Liang Cheng
- Subjects
Gynecology ,medicine.medical_specialty ,Index (economics) ,business.industry ,Urology ,medicine ,Medical physics ,Subject (documents) ,business - Published
- 1999
42. KIT overexpression induces proliferation in astrocytes in an imatinib-responsive manner and associates with proliferation index in gliomas
- Author
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Tea, Blom, Heli, Fox, Alexandre, Angers-Loustau, Karita, Peltonen, Laura, Kerosuo, Kirmo, Wartiovaara, Marika, Linja, Olli A, Jänne, Panu, Kovanen, Hannu, Haapasalo, and Nina N, Nupponen
- Subjects
Antineoplastic Agents ,Apoptosis ,Cell Growth Processes ,Glioma ,Transfection ,Piperazines ,Enzyme Activation ,Mice ,Proto-Oncogene Proteins c-kit ,Pyrimidines ,Matrix Metalloproteinase 9 ,Astrocytes ,Benzamides ,Imatinib Mesylate ,Neoplastic Stem Cells ,Animals ,Humans ,Matrix Metalloproteinase 2 ,RNA, Messenger ,Protein Kinase Inhibitors - Abstract
Activating gene mutations, gene amplifications and overexpressed proteins may be useful as targets for novel therapies. Alterations at chromosome locus 4q12 are associated with gliomas and the region harbors the receptor tyrosine kinase gene KIT, which is frequently amplified in gliomas, and also overexpressed in a subset of gliomas. KIT and its ligand stem cell factor are widely expressed in embryonic and adult mouse brain, and they play a role in many signal transduction pathways involved in cellular proliferation, differentiation and cancer cell metastasis. However, the function of KIT in gliomagenesis or disease progression remains unresolved as well as its role in neural and brain tumor development. In this study, we utilized lentivirus-mediated gene transfer to deliver the KIT gene into mouse astrocytes. The growth properties of KIT overexpressing cells were analyzed using several in vitro functional assays. The effect of receptor tyrosine kinase inhibitor imatinib on astrocyte growth was also investigated. Our results indicate that overexpression of KIT in mouse astrocytes promotes cell proliferation, and the increased proliferation is partly inhibited by imatinib treatment. Furthermore, KIT overexpression induces phenotypic changes in the cells suggesting that KIT may play a role in astrocyte growth regulation.
- Published
- 2008
43. Patterns of PIK3CA alterations in familial colorectal and endometrial carcinoma
- Author
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Päivi Peltomäki, Nina N. Nupponen, Wael M. Abdel-Rahman, Annette Gylling, Ralf Bützow, Marjut Puputti, and Miina Ollikainen
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,Class I Phosphatidylinositol 3-Kinases ,Gene mutation ,medicine.disease_cause ,Proto-Oncogene Proteins p21(ras) ,03 medical and health sciences ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,Germline mutation ,Internal medicine ,Proto-Oncogene Proteins ,medicine ,PTEN ,Humans ,neoplasms ,PI3K/AKT/mTOR pathway ,Polymorphism, Single-Stranded Conformational ,030304 developmental biology ,Aged ,0303 health sciences ,biology ,business.industry ,Reverse Transcriptase Polymerase Chain Reaction ,Endometrial cancer ,Gene Amplification ,PTEN Phosphohydrolase ,Membrane Proteins ,Middle Aged ,medicine.disease ,digestive system diseases ,Lynch syndrome ,3. Good health ,Endometrial Neoplasms ,030220 oncology & carcinogenesis ,Mutation ,biology.protein ,ras Proteins ,Female ,KRAS ,business ,Carcinogenesis ,Colorectal Neoplasms - Abstract
While the phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway is known to be activated in multiple sporadic cancers, the role of this pathway in familial tumors is mostly unknown. We searched for alterations in the catalytic domain of PI3K (PIK3CA), PTEN and KRAS, all of which may contribute to PI3K/AKT pathway activation, in a total of 160-familial colorectal (CRC) and endometrial carcinomas (EC), stratified by the presence vs. absence of germline mutations in DNA mismatch repair (MMR) genes. PIK3CA alterations (consisting of point mutations or low-level amplification, which were mutually exclusive with 1 exception) occurred in 10/70 (14%) of CRCs and 19/90 (21%) of ECs. Within ECs, amplification was significantly associated with the subgroup lacking germline mutations in MMR genes (familial site-specific endometrial cancer) (p = 0.015). Decreased or lost PTEN expression was characteristic of endometrial tumourigenesis (51/81, 63%, in EC compared with 24/62, 39%, in CRC, p = 0.004) and KRAS mutations of colorectal tumourigenesis (19/70, 27% in CRC vs. 9/89, 10%, in EC, p = 0.006) regardless of the MMR gene mutation status. PIK3CA alterations frequently coexisted with PTEN or KRAS changes. Combined with published studies on sporadic tumors, our data broaden the understanding of the role for PI3K pathway genes in human tumorigenesis. © 2007 Wiley-Liss, Inc.
- Published
- 2007
44. Molecular cytogenetic mapping of 24 CEPH YACs and 24 gene-specific large insert probes to chromosome 17
- Author
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Anne Kallioniemi, Ritva Karhu, Nina N. Nupponen, Minna Tanner, Paulina Paavola, Maarit Bärlund, and Olli Kallioniemi
- Subjects
Genetics ,endocrine system ,Genome, Human ,Chromosome Mapping ,Biology ,Cytogenetic map ,Insert (molecular biology) ,Chromosome 17 (human) ,Molecular level ,Gene mapping ,Genetic marker ,Yeasts ,Humans ,DNA Probes ,Chromosomes, Artificial, Yeast ,Molecular Biology ,Gene ,In Situ Hybridization, Fluorescence ,Genetics (clinical) ,Chromosomes, Human, Pair 17 - Abstract
Defining boundaries of chromosomal rearrangements at the molecular level would benefit from landmarks that link the cytogenetic map to physical, genetic, and transcript maps, as well as from large-insert FISH probes for such loci to detect numerical and structural rearrangements in metaphase or interphase cells. Here, we determined the locations of 24 genetically mapped CEPH-Mega YACs along the FLpter scale (fractional length from p-telomere) by quantitative fluorescence in situ hybridization analysis. This generated a set of cytogenetically mapped probes for chromosome 17 with an average spacing of about 5 cM. We then developed large-insert YAC, BAC, PAC, or P1 clones to the following 24 known genes, and determined refined map locations along the same FLpter scale: pter–TP53–TOP3–cen–TNFAIP1–ERBB2–TOP2A–BRCA1–TCF11–NME1–HLF–ZNF147/CLN80–BCL5/MPO/SFRS1–TBX2–PECAM1–DDX5/PRKCA–ICAM2–GH1/PRKAR1A–GRB2–CDK3/FKHL13–qter. Taken together, these 48 cytogenetically mapped large-insert probes provide tools for the molecular analysis of chromosome 17 rearrangements, such as mapping amplification, deletion, and translocation breakpoints in this chromosome, in cancer and other diseases.
- Published
- 1998
45. Amplification of KIT, PDGFRA, VEGFR2, and EGFR in gliomas
- Author
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Hanna Mäenpää, Jorma Isola, Olli Tynninen, Heikki Joensuu, Nina N. Nupponen, Tea Blom, Marjut Puputti, Anders Paetau, and Harri Sihto
- Subjects
Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Chromogenic in situ hybridization ,Gene Expression ,PDGFRA ,03 medical and health sciences ,0302 clinical medicine ,Glioma ,Medicine ,Humans ,neoplasms ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,medicine.diagnostic_test ,business.industry ,Brain Neoplasms ,Gene Amplification ,Amplicon ,medicine.disease ,Vascular Endothelial Growth Factor Receptor-2 ,nervous system diseases ,3. Good health ,ErbB Receptors ,Proto-Oncogene Proteins c-kit ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Immunohistochemistry ,business ,Tyrosine kinase ,Anaplastic astrocytoma ,Fluorescence in situ hybridization - Abstract
Receptor tyrosine kinase aberrations are implicated in the genesis of gliomas. We investigated expression and amplification of KIT, PDGFRA, VEGFR2, and EGFR in 87 gliomas consisting of astrocytomas, anaplastic astrocytomas, oligodendrogliomas, or oligoastrocytomas in tumor samples collected at the time of the diagnosis and in samples of the same tumors at tumor recurrence. Gene amplifications were investigated using either chromogenic in situ hybridization or fluorescence in situ hybridization, and protein expression using immunohistochemistry. In samples collected at glioma diagnosis, KIT and PDGFRA amplifications were more frequent in anaplastic astrocytomas than in astrocytomas, oligodendrogliomas, and oligoastrocytomas [28% versus 5% (P = 0.012) and 33% versus 2% (P = 0.0008), respectively]. VEGFR2 amplifications occurred in 6% to 17% of the gliomas at diagnosis, and EGFR amplifications in 0% to 12%. Amplified KIT was more frequently present in recurrent gliomas than in newly diagnosed gliomas (P = 0.0066). KIT amplification was associated with KIT protein expression and with presence of PDGFRA and EGFR amplifications both at the time of the first glioma diagnosis and at tumor recurrence, and with VEGFR2 amplification at tumor recurrence. Three (4%) primary gliomas and 10 (14%) recurrent gliomas that were evaluable for coamplification of KIT, PDGFRA, and VEGFR2 showed amplification of at least two of these genes; the amplicon contained amplified KIT in all 13 cases. In conclusion, besides glioblastoma, amplified KIT, PDGFRA, and VEGFR may also occur in lower-grade gliomas and in their recurrent tumors. It is currently not known whether specific tyrosine kinase inhibitors are effective in the treatment of such gliomas. (Mol Cancer Res 2006;4(12):927–34)
- Published
- 2006
46. Prostate cancer susceptibility genes: Many studies, many results, no answers
- Author
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Nina N. Nupponen and John D. Carpten
- Published
- 2006
47. Gastrointestinal stromal tumors with KIT exon 11 deletions are associated with poor prognosis
- Author
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Bengt Nilsson, Lars-Gunnar Kindblom, Johanna Andersson, Jeanne M. Meis-Kindblom, Anders Odén, Nina N. Nupponen, Harri Sihto, Heikki Joensuu, Per Bümming, and Bengt Gustavsson
- Subjects
Adult ,Male ,Receptor, Platelet-Derived Growth Factor alpha ,Gastrointestinal Stromal Tumors ,Population ,DNA Mutational Analysis ,Mutation, Missense ,PDGFRA ,Biology ,medicine.disease_cause ,Risk Assessment ,Sensitivity and Specificity ,Cohort Studies ,03 medical and health sciences ,Exon ,0302 clinical medicine ,medicine ,Biomarkers, Tumor ,Missense mutation ,Humans ,Stromal tumor ,education ,030304 developmental biology ,Aged ,Probability ,Retrospective Studies ,0303 health sciences ,education.field_of_study ,Mutation ,Hepatology ,GiST ,Gastroenterology ,Exons ,Middle Aged ,Prognosis ,digestive system diseases ,3. Good health ,Gene Expression Regulation, Neoplastic ,Survival Rate ,genomic DNA ,Proto-Oncogene Proteins c-kit ,030220 oncology & carcinogenesis ,Cancer research ,Female ,Gene Deletion - Abstract
Background & Aims: Gain-of-function mutations in the KIT receptor tyrosine kinase gene and rare mutations in the platelet-derived growth factor receptor α ( PDGFRA ) gene are important events in gastrointestinal stromal tumor (GIST) development. Different mutations are reportedly associated with distinctive phenotypes and possibly clinical behavior. We investigated the correlation among mutation type, phenotype, and clinical course in a preimatinib, population-based series of GIST with long-term follow-up. Methods: Genomic DNA from 177 GIST patients was analyzed for KIT exons 9, 11, 13, and 17 and PDGFRA exons 12 and 18 mutations using denaturating high-performance liquid chromatography and bidirectional sequencing. Results:KIT exon 11 mutations were detected in 101 of 177 GIST (61 deletions, 23 missense mutations, and 17 duplications); wild-type (WT) KIT and PDGFRA were detected in 63; KIT exon 9 and exon 17 mutations in 6 and 1, respectively; and PDGFRA exons 12 and 18 mutations in 3 each. GIST >5 cm vs GIST ≤1 cm had mutations in 73% and 33%, respectively. KIT exon 11 deletions were significantly associated with a higher proportion of high risk or overtly malignant groups compared with WT GIST. KIT exon 11 deletions adversely affected outcome. KIT exon 11 duplications and exon 9 mutations were found exclusively in gastric and small intestinal GIST, respectively. Conclusions: KIT exon 11 deletion is an independent adverse prognostic factor in patients with GIST.
- Published
- 2005
48. Primary cutaneous T-cell lymphomas show a deletion or translocation affecting NAV3, the human UNC-53 homologue
- Author
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Bogusław Nedoszytko, Vesna Blazevic, Nina N. Nupponen, Hanna Nevala, Sonja Hahtola, Suvi Päivinen, Ying Zhou, Annamari Ranki, Pärt Peterson, Sanna Syrjä, Jadwiga Roszkiewicz, Ritva Karhu, Maria Pesonen, Inge Krebs, Kalle Saksela, Leena Karenko, Marketta Kähkönen, Harri Sihto, Tapio Visakorpi, Soili Kytölä, and Annemarie Poustka
- Subjects
Cancer Research ,Monosomy ,Pathology ,medicine.medical_specialty ,Skin Neoplasms ,Tumor suppressor gene ,Chromosomal translocation ,Nerve Tissue Proteins ,Biology ,Translocation, Genetic ,medicine ,Humans ,Sezary Syndrome ,Gene Silencing ,Interphase ,Chromosome 12 ,Alleles ,In Situ Hybridization, Fluorescence ,Chromosome Aberrations ,Mycosis fungoides ,Chromosomes, Human, Pair 12 ,Cytogenetics ,Chromosome Mapping ,Membrane Proteins ,Chromosome Breakage ,medicine.disease ,Lymphoma ,Lymphoma, T-Cell, Cutaneous ,Oncology ,Cancer research ,Interleukin-2 ,Haploinsufficiency ,Gene Deletion - Abstract
Multicolor fluorescent in situ hybridization (FISH) was used to identify acquired chromosomal aberrations in 12 patients with mycosis fungoides or Sézary syndrome, the most common forms of primary cutaneous T-cell lymphoma (CTCL). The most frequently affected chromosome was 12, which showed clonal deletions or translocations with a break point in 12q21 or 12q22 in five of seven consecutive Sézary syndrome patients and a clonal monosomy in the sixth patient. The break point of a balanced translocation t(12;18)(q21;q21.2), mapped in the minimal common region of two deletions, fine mapped to 12q2. By locus-specific FISH, the translocation disrupted one gene, NAV3 (POMFIL1), a human homologue of unc-53 in Caenorhabditis elegans. A missense mutation in the remaining NAV3 allele was found in one of six cases with a deletion or translocation. With locus-specific FISH, NAV3 deletions were found in the skin lesions of four of eight (50%) patients with early mycosis fungoides (stages IA-IIA) and in the skin or lymph node of 11 of 13 (85%) patients with advanced mycosis fungoides or Sézary syndrome. Preliminary functional studies with lentiviral small interfering RNA-based NAV3 silencing in Jurkat cells and in primary lymphocytes showed enhanced interleukin 2 expression (but not CD25 expression). Thus, NAV3 may contribute to the growth, differentiation, and apoptosis of CTCL cells as well as to the skewing from Th1-type to Th2-type phenotype during disease progression. NAV3, a novel putative haploinsufficient tumor suppressor gene, is disrupted in most cases of the commonest types of CTCL and may thus provide a new diagnostic tool.
- Published
- 2005
49. Mutations in two short noncoding mononucleotide repeats in most microsatellite-unstable colorectal cancers
- Author
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Virpi Launonen, Heikki Järvinen, Tuija Hienonen, Pia Alhopuro, Nina N. Nupponen, Heli J. Lehtonen, Bert Vogelstein, Auli Karhu, Susa Enholm, Reijo Salovaara, Diego Arango, Jukka-Pekka Mecklin, Lauri A. Aaltonen, Riitta Koistinen, Thomas D. Barber, Rainer Lehtonen, and Heli Sammalkorpi
- Subjects
Genome instability ,Cancer Research ,DNA Repair ,DNA repair ,Base Pair Mismatch ,Biology ,medicine.disease_cause ,Seminal Vesicle Secretory Proteins ,Genomic Instability ,Frameshift mutation ,03 medical and health sciences ,0302 clinical medicine ,Cell Line, Tumor ,medicine ,Humans ,RNA, Messenger ,Mutation frequency ,Frameshift Mutation ,Alleles ,030304 developmental biology ,Genetics ,0303 health sciences ,Mutation ,Base Sequence ,Microsatellite instability ,DNA, Neoplasm ,medicine.disease ,Introns ,3. Good health ,Semenogelin I ,Oncology ,030220 oncology & carcinogenesis ,DNA mismatch repair ,Colorectal Neoplasms ,Microsatellite Repeats - Abstract
DNA mismatch repair (MMR)–deficient cells typically accumulate mutations in short repetitive DNA tracts. This microsatellite instability (MSI) facilitates malignant transformation when affecting genes with growth-related and caretaker functions. To date, several putative MSI target genes have been proposed mainly based on high mutation frequency within their coding regions. However, some intronic repeat mutations have also been suggested to associate with MSI tumorigenesis, indicating the need for additional analyses on noncoding repeats. Here we have analyzed an intronic T9 repeat of semenogelin I (SEMG1) and report mutation frequencies of 51% (75 of 146) and 62% (8 of 13) in MMR-deficient primary colorectal cancers and cell lines, respectively. The putative effect of the SEMG1 mutations was assessed by RNA and protein level analyses, but no differences were detected between colorectal cancer cell lines with different SEMG1 status. Subsequently, the general background mutation frequency of MSI colorectal cancers was assessed by screening for intergenic T9 repeat alterations. One of 10 examined repeats was mutated in 70% (102 of 145) of the colorectal cancers evaluated. The frequencies observed here are notably higher than previously published in noncoding repeats shorter than 10 bp in MMR-deficient primary tumors. Our results indicate that high mutation frequencies, similar or higher than those observed in proposed and approved target genes, can be detected in repeat tracts of MSI tumors without any apparent selection pressure. These data call for urgent and thorough large-scale evaluation of mutation frequencies in neutral short repetitive sequences in MMR-deficient tumors.
- Published
- 2005
50. KIT and platelet-derived growth factor receptor alpha tyrosine kinase gene mutations and KIT amplifications in human solid tumors
- Author
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Kaarle Franssila, Harri Sihto, Olli Tynninen, Nina N. Nupponen, Minna Tanner, Maarit Sarlomo-Rikala, Leif C. Andersson, and Heikki Joensuu
- Subjects
Male ,Teratocarcinoma ,Cancer Research ,Receptor, Platelet-Derived Growth Factor alpha ,Platelet-Derived Growth Factor Receptor Alpha ,PDGFRA ,Gene mutation ,Polymerase Chain Reaction ,Growth factor receptor ,Testicular Neoplasms ,medicine ,Humans ,Carcinoma, Small Cell ,In Situ Hybridization, Fluorescence ,Gastrointestinal Neoplasms ,Oncogene Proteins ,biology ,CD117 ,Gene Amplification ,Cancer ,Exons ,Protein-Tyrosine Kinases ,medicine.disease ,Aneuploidy ,Molecular biology ,Immunohistochemistry ,digestive system diseases ,Proto-Oncogene Proteins c-kit ,Imatinib mesylate ,Oncology ,Mutation ,biology.protein ,Cancer research ,Chromosomes, Human, Pair 4 ,Tyrosine kinase - Abstract
Purpose Mutated KIT and platelet-derived growth factor receptor alpha (PDGFRα) tyrosine kinases are the principal targets for imatinib mesylate in the treatment of gastrointestinal stromal tumors (GISTs). The frequency of activating KIT and PDGFRA gene mutations in most other histologic types of human cancer is not known. Materials and Methods KIT exons 9, 11, 13, and 17 and PDGFRA exons 11 and 17 of 334 human cancers were screened for mutations using sensitive denaturing high-performance liquid chromatography (DHPLC). In addition, all KIT exons from 9 to 21 of 115 tumors were screened. Thirty-two histologic tumor types were examined. Samples with abnormal findings in DHLPC were sequenced. Immunostaining for the KIT protein (CD117) was performed in 322 (96.4%) of the 334 cases. Results Of the 3,039 exons screened, only 17 had mutation. All 17 cases with either mutated KIT (n = 15) or PDGFRA (n = 2) were histologically GIST tumors, whereas none of the other histologic types of cancer (n = 316) harbored KIT or PDGFRA mutation. KIT immunostaining was rarely positive except in GISTs (18 of 18), small-cell lung cancer (10 of 30; 33%), and testicular teratocarcinoma (four of 17; 24%). Wild-type KIT gene amplification or chromosome 4 aneuploidy was common (seven of 12) in non-GIST tumors with strong KIT protein expression when studied with fluorescence in situ hybridization. Conclusion Despite frequent KIT protein expression in some tumor types, KIT and PDGFRA gene mutations are uncommon in most human cancers. Cancer KIT expression is frequently associated with multiple copies of the wild-type KIT gene.
- Published
- 2004
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