18 results on '"Mahboubeh Papari-Zareei"'
Search Results
2. Cell-autonomous immune gene expression is repressed in pulmonary neuroendocrine cells and small cell lung cancer
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Jane E. Johnson, Hongyu Liu, John D. Minna, Guanghua Xiao, Benjamin J. Drapkin, Junya Fujimoto, Adi F. Gazdar, Yang Xie, Ling Cai, Kenneth E. Huffman, Yu An Zhang, Tao Wang, Gaoxiang Jia, Ignacio I. Wistuba, Adeeb Alomar, Victor Stastny, Fang Huang, Jun Chen, Karine Pozo, Esra A. Akbay, Dhruba Deb, Luc Girard, Yongwen Li, Ralph J. DeBerardinis, Lin Yang, Mahboubeh Papari-Zareei, Chendong Yang, David S. Shames, Wei Zou, and Christin S. Kuo
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0301 basic medicine ,Lung Neoplasms ,QH301-705.5 ,Genes, MHC Class I ,Medicine (miscellaneous) ,Antineoplastic Agents ,Article ,Small-cell lung cancer ,General Biochemistry, Genetics and Molecular Biology ,Immunophenotyping ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Cell Line, Tumor ,MHC class I ,Biomarkers, Tumor ,Tumor Microenvironment ,medicine ,Animals ,Humans ,Neoplasm ,Cell Lineage ,Biology (General) ,Data mining ,neoplasms ,Regulation of gene expression ,Tumor microenvironment ,biology ,Gene Expression Profiling ,medicine.disease ,Small Cell Lung Carcinoma ,Xenograft Model Antitumor Assays ,Phenotype ,humanities ,respiratory tract diseases ,Gene Expression Regulation, Neoplastic ,Gene expression profiling ,Neuroendocrine Tumors ,030104 developmental biology ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,biology.protein ,Cancer research ,Transcriptome ,General Agricultural and Biological Sciences - Abstract
Small cell lung cancer (SCLC) is classified as a high-grade neuroendocrine (NE) tumor, but a subset of SCLC has been termed “variant” due to the loss of NE characteristics. In this study, we computed NE scores for patient-derived SCLC cell lines and xenografts, as well as human tumors. We aligned NE properties with transcription factor-defined molecular subtypes. Then we investigated the different immune phenotypes associated with high and low NE scores. We found repression of immune response genes as a shared feature between classic SCLC and pulmonary neuroendocrine cells of the healthy lung. With loss of NE fate, variant SCLC tumors regain cell-autonomous immune gene expression and exhibit higher tumor-immune interactions. Pan-cancer analysis revealed this NE lineage-specific immune phenotype in other cancers. Additionally, we observed MHC I re-expression in SCLC upon development of chemoresistance. These findings may help guide the design of treatment regimens in SCLC., Ling Cai et al. used transcriptomic profiling data of healthy lung, patient-derived small cell lung cancer cell lines, xenografts, and primary tumors to examine a link between neuroendocrine (NE) signatures and immune gene expression. Their findings suggest that cell-autonomous immune gene repression is a shared feature between healthy and tumor cells of NE lineage and may influence tumor-immune cell interaction and response to immunotherapy.
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- 2021
3. Relationship Between Neuroendocrine and Immune Gene Expression in Small Cell Lung Cancer
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Ling Cai, Hongyu Liu, fang huang, Junya Fujimoto, Luc Girard, Jun Chen, Yongwen Li, Yu-an Zhang, Dhruba Deb, Victor Stastny, Christin Kuo, Gaoxiang Jia, Chendong Yang, Wei Zou, Adeeb Alomar, Kenneth Huffman, Mahboubeh Papari-Zareei, Lin Yang, Benjamin Drapkin, Esra Akbay, David Shames, Ignacio Wistuba, Tao Wang, Guanghua Xiao, Ralph DeBerardinis, John D. Minna, Yang Xie, and Adi Gazdar
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neoplasms ,humanities ,respiratory tract diseases - Abstract
Small cell lung cancer (SCLC) is classified as a high-grade neuroendocrine (NE) tumor, but a subset of SCLC has been termed “variant” due to the loss of NE characteristics. In this study, we computed NE scores for patient-derived SCLC cell lines and xenografts, as well as human tumors. We aligned NE properties with transcription factor-defined molecular subtypes. Then we investigated the different immune phenotypes associated with high and low NE scores. We found repression of immune response genes as a shared feature between classic SCLC and pulmonary neuroendocrine cells of the healthy lung. With loss of NE fate, variant SCLC tumors regain cell-autonomous immune gene expression and exhibit higher tumor-immune interactions. Pan-cancer analysis revealed this NE lineage-specific immune phenotype in other cancers. Additionally, we observed MHC I re-expression in SCLC upon development of chemoresistance. These findings provide a new framework to guide design of treatment regimens in SCLC.
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- 2020
4. Relationship Between Neuroendocrine and Immune Gene Expression in Small Cell Lung Cancer
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Jun Chen, Gaoxiang Jia, Yuan Zhang, Yang Xie, Kenneth E. Huffman, Yongwen Li, Adi F. Gazdar, Guanghua Xiao, Adeeb Alomar, Victor Stastny, Chendong Yang, Junya Fujimoto, David S. Shames, John D. Minna, Fang Huang, Dhruba Deb, Luc Girard, Ralph J. DeBerardinis, Tao Wang, Benjamin J. Drapkin, Hongyu Liu, Ling Cai, Christin S. Kuo, Ignacio I. Wistuba, Esra A. Akbay, Lin Yang, Mahboubeh Papari-Zareei, and Wei Zou
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Lung ,Biology ,Phenotype ,humanities ,respiratory tract diseases ,Immune system ,medicine.anatomical_structure ,Transcription (biology) ,Cell culture ,MHC class I ,Cancer research ,medicine ,biology.protein ,neoplasms ,Psychological repression ,Immune gene - Abstract
Small cell lung cancer (SCLC) is classified as a high-grade neuroendocrine (NE) tumor, but a subset of SCLC has been termed “variant” due to the loss of NE characteristics. In this study, we computed NE scores for patient-derived SCLC cell lines and xenografts, as well as human tumors. We aligned NE properties with transcription factor-defined molecular subtypes. Then we investigated the different immune phenotypes associated with high and low NE scores. We found repression of immune response genes as a shared feature between classic SCLC and pulmonary neuroendocrine cells of the healthy lung. With loss of NE fate, variant SCLC tumors regain cell-autonomous immune gene expression and exhibit higher tumor-immune interactions. Pan-cancer analysis revealed this NE lineage-specific immune phenotype in other cancers. Additionally, we observed MHC I re-expression in SCLC upon development of chemoresistance. These findings provide a new framework to guide design of treatment regimens in SCLC.
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- 2020
5. Small cell lung cancer tumors and preclinical models display heterogeneity of neuroendocrine phenotypes
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Wei Zhang, Luc Girard, Hans K. Ghayee, Victor Stastny, Adi F. Gazdar, T. Haruki, Mahboubeh Papari-Zareei, Yu An Zhang, Karel Pacak, Trudy G. Oliver, and John D. Minna
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0301 basic medicine ,Oncogene ,business.industry ,Original Article on Small Cell Lung Cancer: New Models, Markers and Beyond ,Neuroendocrine tumors ,medicine.disease ,Phenotype ,respiratory tract diseases ,03 medical and health sciences ,ASCL1 ,030104 developmental biology ,Oncology ,Cell culture ,NEUROD1 ,Cancer research ,Medicine ,Epithelial–mesenchymal transition ,business ,Lung cancer - Abstract
Background Small cell lung cancer (SCLC) is a deadly, high grade neuroendocrine (NE) tumor without recognized morphologic heterogeneity. However, over 30 years ago we described a SCLC subtype with "variant" morphology which did not express some NE markers and exhibited more aggressive growth. Methods To quantitate NE properties of SCLCs, we developed a 50-gene expression-based NE score that could be applied to human SCLC tumors and cell lines, and genetically engineered mouse (GEM) models. We identified high and low NE subtypes of SCLC in all of our sample types, and characterized their properties. Results We found that 16% of human SCLC tumors and 10% of SCLC cell lines were of the low NE subtype, as well as cell lines from the GEM model. High NE SCLC lines grew as non-adherent floating aggregates or spheroids while Low NE lines had morphologic features of the variant subtype and grew as loosely attached cells. While the high NE subtype expressed one of the NE lineage master transcription factors ASCL1 or NEUROD1, together with NKX2-1, the entire range of NE markers, and lacked expression of the neuronal and NE repressor REST, the low NE subtype had lost expression of most NE markers, ASCL1, NEUROD1 and NKX2-1 and expressed REST. The low NE subtype had undergone epithelial mesenchymal transition (EMT) and had activated the Notch, Hippo and TGFβ pathways and MYC oncogene . Importantly, the high and low NE group of SCLC lines had similar gene expression profiles as their SCLC tumor counterparts. Conclusions SCLC tumors and cell lines can exhibit distinct inter-tumor heterogeneity with respect to expression of NE features. Loss of NE expression results in major alterations in morphology, growth characteristics, and molecular properties. These findings have major clinical implications as the two subtypes are predicted to have very different responses to targeted therapies.
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- 2018
6. Phenotypic variation in a Chinese family with 46,XY and 46,XX 17α-hydroxylase deficiency
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Chandran Manju, Yong Mong Bee, Mahboubeh Papari-Zareei, and Richard J. Auchus
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Adult ,Proband ,China ,endocrine system ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,DNA Mutational Analysis ,Biology ,Compound heterozygosity ,Young Adult ,Exon ,Endocrinology ,Asian People ,Internal medicine ,medicine ,Humans ,Missense mutation ,Gene ,Genetics ,Adrenal Hyperplasia, Congenital ,Steroid 17-alpha-Hydroxylase ,Obstetrics and Gynecology ,Sexual Infantilism ,Phenotype ,CYP17A1 ,Mutation ,Female ,Amenorrhea ,medicine.symptom - Abstract
17α-hydroxylase deficiency is a rare autosomal recessive disorder characterized by sexual infantilism, amenorrhea, hypertension and hypokalemia, which is caused by mutations in the CYP17A1 gene. To date, more than 50 mutations in this gene have been described.The clinical features and biochemical data of a pair of 46,XY and 46,XX Chinese siblings with 17α-hydroxylase deficiency from Singapore were studied. Direct DNA sequence analysis of the CYP17A1 gene was performed.There was significant phenotypic variation between the siblings. The proband (46,XY) presented classically with sexual infantilism, amenorrhea and hypertension. The younger sibling (46,XX) also presented with amenorrhea, but she had breast development and absence of hypokalemic hypertension. The same compound heterozygous mutations in CYP17A1 gene were identified in both patients. A missense mutation (P409R) was detected in exon 7, and a 9-bp deletion (D487-S488-F489del) was detected in exon 8.We confirmed the diagnosis of 17α-hydroxylase deficiency in these two patients. Both P409R and D487-S488-F489del have been described previously and are widely propagated in the Chinese population in East and Southeast Asia. We propose that the phenotypic expression of affected individuals with 17α-hydroxylase deficiency is karyotype-dependent, with individuals having the 46,XX karyotype having less pronounced clinical symptoms.
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- 2011
7. Abstract 2664: Identification of two small molecules with small cell lung cancer growth inhibition response profiles different from etoposide/topotecan
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Michael G. Roth, John B. MacMillan, John D. Minna, Kimberley Avila, Gazdar Adi, Elizabeth A. McMillan, Luc Girard, Bruce A. Posner, Michael Peyton, Victor Stastny, Michael A. White, Paul Yenerall, T. Haruki, Mahboubeh Papari-Zareei, Kenneth E. Huffman, and Allison A. Mootz
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Cancer Research ,Etoposide/Topotecan ,business.industry ,Cancer ,medicine.disease ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Oncology ,chemistry ,030220 oncology & carcinogenesis ,Cancer research ,Medicine ,030211 gastroenterology & hepatology ,Topotecan ,Non small cell ,Growth inhibition ,business ,Survival rate ,Etoposide ,medicine.drug - Abstract
Small Cell Lung Cancer (SCLC) is an aggressive, highly metastatic cancer with an overall 5-year survival rate of Citation Format: Allison A. Mootz, Michael Peyton, Paul Yenerall, Kimberley Avila, Kenneth Huffman, Tomohiro Haruki, Mahboubeh Papari-Zareei, Victor Stastny, Luc Girard, Elizabeth McMillan, Michael Roth, John MacMillan, Bruce Posner, Michael White, Gazdar Adi, John Minna. Identification of two small molecules with small cell lung cancer growth inhibition response profiles different from etoposide/topotecan [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2664.
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- 2018
8. A Novel Point Mutation in P450c17 (CYP17) Causing Combined 17α-Hydroxylase/17,20-Lyase Deficiency
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J. H. Shepherd, Martin Gore, John Achermann, T. Ahmad, Gill Rumsby, John P Monson, A. M. Brooke, Lin Lin, Norman F. Taylor, Richard J. Auchus, and Mahboubeh Papari-Zareei
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Models, Molecular ,medicine.medical_specialty ,Adolescent ,Arginine ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,Mutation, Missense ,Context (language use) ,Biology ,medicine.disease_cause ,Biochemistry ,Endocrinology ,Internal medicine ,medicine ,Humans ,Point Mutation ,Missense mutation ,Congenital adrenal hyperplasia ,Mutation ,Adrenal Hyperplasia, Congenital ,Point mutation ,Biochemistry (medical) ,Steroid 17-alpha-Hydroxylase ,medicine.disease ,Lyase ,Mutation testing ,Female - Abstract
Combined 17alpha-hydroxylase/17,20-lyase deficiency is a rare cause of congenital adrenal hyperplasia and hypogonadism. Novel single amino acid changes in P450c17 provide potentially important insights into key structural domains for enzyme function.We report a novel missense mutation in P450c17 in a 17-yr-old female presenting with a malignant mixed germ cell tumor with yolk sac elements who demonstrated clinical and biochemical features of combined 17alpha-hydroxylase/17,20-lyase deficiency.Quantitative urinary steroid analysis was performed by high resolution gas chromatography. All eight coding exons of CYP17 were PCR amplified and sequenced. The position of arginine at codon 96 was modeled using the CYP17 structure 2c17 (www.rcsb.org). The CYP17 genes were subcloned into pcDNA3, expressed in HEK-293 cells, and chromatographed.17alpha-Hydroxylase deficiency was confirmed by marked reductions in urinary and serum cortisol, androgens, and estradiol. Mutational analysis revealed a novel homozygous R96Q missense mutation in P450c17, affecting an amino acid in a key substrate-binding region of the enzyme, leading to complete inactivity.The description of a second missense mutation at codon 96 (R96W and R96Q) in the substrate-binding region of P450c17 provides strong evidence for the key role of this amino acid in 17alpha-hydroxylase/17,20-lyase function. An association between a malignant germ cell tumor and 17alpha-hydroxylase deficiency has not been reported previously, although the presence of gonadoblastoma in the ovary of a patient with this condition has recently been described.
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- 2006
9. Arginine 276 Controls the Directional Preference of AKR1C9 (Rat Liver 3α-Hydroxysteroid Dehydrogenase) in Human Embryonic Kidney 293 Cells
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Mahboubeh Papari-Zareei, Richard J. Auchus, and Andrew Brandmaier
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Androstenediol ,medicine.medical_specialty ,Arginine ,Dehydrogenase ,Reductase ,Biology ,Kidney ,Structure-Activity Relationship ,Endocrinology ,Oxidoreductase ,Internal medicine ,medicine ,Animals ,Humans ,Cells, Cultured ,chemistry.chemical_classification ,HEK 293 cells ,Dihydrotestosterone ,Rats ,Enzyme ,chemistry ,NAD+ kinase ,Oxidoreductases ,NADP ,medicine.drug - Abstract
Rat liver AKR1C9 is the best-studied 3alpha-hydroxysteroid dehydrogenase (3alphaHSD) of the aldo-keto reductase superfamily. The physiologic function of AKR1C9 is to catalyze the reduction of 5alpha-androstane-17beta-ol-3-one (dihydrotestosterone) to 5alpha-androstane-3alpha,17beta-diol (androstanediol) rather than the reverse reaction, and all of the known AKR1C enzymes with 3alphaHSD activity also preferentially catalyze dihydrotestosterone reduction in intact cells. Because the utilization of pyridine-nucleotide cofactors NAD(P)(H) primarily governs the directional preference of HSD enzymes in intact cells, and because R276 participates in NADP(H) binding, we hypothesized that mutation of R276 would alter directional preference in intact cells. To test this model, we constructed stable lines of human embryonic kidney 293 cells expressing wild-type AKR1C9 and mutations R276M, R276G, and R276E. Mutations R276M and R276G retained reductive preference with slightly reduced magnitude compared with wild-type AKR1C9. NADPH depletion by glucose deprivation minimally altered the equilibrium steroid distribution for wild-type AKR1C9 but further reduced the reductive preference of mutations R276M and R276G. Mutation R276E, in contrast, showed an oxidative preference under all conditions. The intrinsic rates of the reductive and oxidative reactions for all four enzymes were similar at the functional equilibrium states. We conclude the R276 maximizes the reductive preference of AKR1C9 in intact cells and maintains this strong preference despite NADPH depletion; mutation R276E reverses the directional preference.
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- 2006
10. Dihydrotestosterone synthesis bypasses testosterone to drive castration-resistant prostate cancer
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Rui Li, Nima Sharifi, Lori M Watumull, Yan D. Zhao, Richard J. Auchus, Mahboubeh Papari-Zareei, and Kai Hsiung Chang
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Male ,medicine.medical_specialty ,medicine.drug_class ,Transplantation, Heterologous ,Biology ,urologic and male genital diseases ,Androgen deprivation therapy ,chemistry.chemical_compound ,Prostate cancer ,Mice ,3-Oxo-5-alpha-Steroid 4-Dehydrogenase ,Internal medicine ,medicine ,Animals ,Humans ,Testosterone ,Castration ,Multidisciplinary ,Abiraterone acetate ,Membrane Proteins ,Prostatic Neoplasms ,Dihydrotestosterone ,Biological Sciences ,medicine.disease ,Androgen ,SRD5A1 ,Endocrinology ,chemistry ,SRD5A2 ,Disease Progression ,Metabolic Networks and Pathways ,medicine.drug - Abstract
In the majority of cases, advanced prostate cancer responds initially to androgen deprivation therapy by depletion of gonadal testosterone. The response is usually transient, and metastatic tumors almost invariably eventually progress as castration-resistant prostate cancer (CRPC). The development of CRPC is dependent upon the intratumoral generation of the potent androgen, dihydrotestosterone (DHT), from adrenal precursor steroids. Progression to CRPC is accompanied by increased expression of steroid-5α-reductase isoenzyme-1 (SRD5A1) over SRD5A2, which is otherwise the dominant isoenzyme expressed in the prostate. DHT synthesis in CRPC is widely assumed to require 5α-reduction of testosterone as the obligate precursor, and the increased expression of SRD5A1 is thought to reflect its role in converting testosterone to DHT. Here, we show that the dominant route of DHT synthesis in CRPC bypasses testosterone, and instead requires 5α-reduction of androstenedione by SRD5A1 to 5α-androstanedione, which is then converted to DHT. This alternative pathway is operational and dominant in both human CRPC cell lines and fresh tissue obtained from human tumor metastases. Moreover, CRPC growth in mouse xenograft models is dependent upon this pathway, as well as expression of SRD5A1. These findings reframe the fundamental metabolic pathway that drives CRPC progression, and shed light on the development of new therapeutic strategies.
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- 2011
11. Combined 17α-hydroxylase/17,20-lyase deficiency due to p.R96W mutation in the CYP17 gene in a Brazilian patient
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Claudio E. Kater, Richard J. Auchus, Mahboubeh Papari-Zareei, Fabiola Costenaro, Ticiana da Costa Rodrigues, and Mauro Antonio Czepielewski
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Hipogonadismo ,Pediatrics ,medicine.medical_specialty ,Adolescent ,Endocrinology, Diabetes and Metabolism ,Disease ,Esteróide 17-alfa-hidroxilase ,medicine.disease_cause ,Exon ,Genotype ,medicine ,Humans ,Congenital adrenal hyperplasia ,Gene ,Mutation ,Adrenal Hyperplasia, Congenital ,business.industry ,Hypogonadism ,Homozygote ,Steroid 17-alpha-Hydroxylase ,General Medicine ,medicine.disease ,Hypokalemia ,Hiperplasia suprarrenal congênita ,Hypertension ,Female ,Differential diagnosis ,medicine.symptom ,business ,Brazil ,Hipertensão - Abstract
A hiperplasia adrenal congênita (HAC), em razão da deficiência de 17α -hidroxilase/17,20-liase, é uma doença autossômica recessiva rara e a segunda causa mais comum de HAC no Brasil. Descrevemos o caso de um paciente brasileiro portador da deficiência 17α -hidroxilase/17,20- liase (CYP17) em homozigose para a mutação p.R96W no éxon 1 do gene da CYP17A1, uma mutação incomum entre os casos brasileiros descritos com essa forma de HAC. Esse paciente, criado como um indivíduo normal do sexo feminino, procurou atendimento por ausência de sinais puberais e amenorreia primária aos 16 anos de idade. Durante a avaliação, constataram-se um cariótipo 46,XY e a presença de hipertensão e hipocalemia. Enfatizamos o reconhecimento da deficiência da CYP17 dentre os possíveis diagnósticos em um paciente jovem com hipogonadismo hipergonadotrófico e hipertensão, os quais necessitam de tratamento particularizado para ambas as situações. Congenital adrenal hyperplasia (CAH) resulting from 17α -hydroxylase/17,20-lyase deficiency is a rare autosomal recessive disease and the second most common form of CAH in Brazil. We describe the case of a Brazilian patient with CYP17 deficiency (17α -hydroxylase/17,20-lyase deficiency) caused by a homozygous p.R96W mutation on exon 1 of the CYP17 gene, an unusual genotype in Brazilian patients with this form of CAH. The patient, raised as a normal female, sought medical care for lack of pubertal signs and primary amenorrhea at the age of 16 years. At evaluation, the presence of a 46,XY karyotype, hypertension and hypokalemia were observed. We emphasize the recognition of CYP17 deficiency in the differential diagnosis of cases of hypergonadotrophic hypogonadism and hypertension in young patients who need specific treatment for both situations.
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- 2010
12. 3beta-hydroxysteroid dehydrogenase is a possible pharmacological target in the treatment of castration-resistant prostate cancer
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Kristen Evaul, Rui Li, Mahboubeh Papari-Zareei, Richard J. Auchus, and Nima Sharifi
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Male ,medicine.medical_specialty ,17-Hydroxysteroid Dehydrogenases ,Androstenediol ,Drug Evaluation, Preclinical ,Dehydroepiandrosterone ,Antineoplastic Agents ,Biology ,urologic and male genital diseases ,TMPRSS2 ,Models, Biological ,Androgen deprivation therapy ,chemistry.chemical_compound ,Prostate cancer ,Endocrinology ,Drug Delivery Systems ,Internal medicine ,Cell Line, Tumor ,medicine ,Humans ,Orchiectomy ,Treatment Failure ,Enzyme Inhibitors ,Cell Nucleus ,Dose-Response Relationship, Drug ,Carcinoma ,Prostatic Neoplasms ,Dihydrotestosterone ,medicine.disease ,Androgen receptor ,chemistry ,Receptors, Androgen ,medicine.drug - Abstract
Prostate cancer usually responds to androgen deprivation therapy, although the response in metastatic disease is almost always transient and tumors eventually progress as castration-resistant prostate cancer (CRPC). CRPC continues to be driven by testosterone or dihydrotestosterone from intratumoral metabolism of 19-carbon adrenal steroids from circulation, and/or de novo intratumoral steroidogenesis. Both mechanisms require 3beta-hydroxysteroid dehydrogenase (3betaHSD) metabolism of Delta(5)-steroids, including dehydroepiandrosterone (DHEA) and Delta(5)-androstenediol (A5diol), to testosterone. In contrast, reports that DHEA and A5diol directly activate the androgen receptor (AR) suggest that 3betaHSD metabolism is not required and that 3betaHSD inhibitors would be ineffective in the treatment of CRPC. We hypothesized that activation of AR in prostate cancer by DHEA and A5diol requires their conversion via 3betaHSD to androstenedione and testosterone, respectively. Here, we show that DHEA and A5diol induce AR chromatin occupancy and AR-regulated genes. Furthermore, we show that Delta(5)-androgens undergo 3beta-dehydrogenation in prostate cancer and that induction of AR nuclear translocation, AR chromatin occupancy, transcription of PSA, TMPRSS2, and FKBP5, as well as cell proliferation by DHEA and A5diol, are all blocked by inhibitors of 3betaHSD. These findings demonstrate that DHEA and A5diol must be metabolized by 3betaHSD to activate AR in these models of CRPC. Furthermore, this work suggests that 3betaHSD may be exploited as a pharmacologic target in the treatment of CRPC.
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- 2010
13. Biochemical factors governing the steady-state estrone/estradiol ratios catalyzed by human 17beta-hydroxysteroid dehydrogenases types 1 and 2 in HEK-293 cells
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Sharareh Akbar, Siayareh Rambally, Richard J. Auchus, Daniel Sherbet, Dario Mizrachi, Mahboubeh Papari-Zareei, and Oleg Guryev
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medicine.medical_specialty ,17-Hydroxysteroid Dehydrogenases ,Estrone ,Molecular Sequence Data ,Estradiol Dehydrogenases ,Dehydrogenase ,Nicotinamide adenine dinucleotide ,Kidney ,Redox ,Cofactor ,Article ,Cell Line ,chemistry.chemical_compound ,Endocrinology ,Oxidoreductase ,Internal medicine ,medicine ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,chemistry.chemical_classification ,biology ,Estradiol ,Kinetics ,chemistry ,Biochemistry ,Amino Acid Substitution ,biology.protein ,NADPH binding ,NAD+ kinase ,Sequence Alignment ,NADP - Abstract
Human 17β-hydroxysteroid dehydrogenase types 1 and 2 (17βHSD1 and 17βHSD2) regulate estrogen potency by catalyzing the interconversion of estrone (E1) and estradiol (E2) using nicotinamide adenine dinucleotide (phosphate) cofactors NAD(P)(H). In intact cells, 17βHSD1 and 17βHSD2 establish pseudo-equilibria favoring E1 reduction or E2 oxidation, respectively. The vulnerability of these equilibrium steroid distributions to mutations and to altered intracellular cofactor abundance and redox state, however, is not known. We demonstrate that the equilibrium E2/E1 ratio achieved by 17βHSD1 in intact HEK-293 cell lines is progressively reduced from 94:6 to 10:90 after mutagenesis of R38, which interacts with the 2′-phosphate of NADP(H), and by glucose deprivation, which lowers the NADPH/NADP+ ratio. The shift to E2 oxidation parallels changes in apparent Km values for purified 17βHSD1 proteins to favor NAD(H) over NADP(H). In contrast, mutagenesis of E116 (corresponding to R38 in 17βHSD1) and changes in intracellular cofactor ratios do not alter the greater than 90:10 E1/E2 ratio catalyzed by 17βHSD2, and these mutations lower the apparent Km of recombinant 17βHSD2 for NADP(H) only less than 3-fold. We conclude that the equilibrium E1/E2 ratio maintained by human 17βHSD1 in intact cells is governed by NADPH saturation, which is strongly dependent on both R38 and high intracellular NADPH/NADP+ ratios. In contrast, the preference of 17βHSD2 for E2 oxidation strongly resists alteration by genetic and metabolic manipulations. These findings suggest that additional structural features, beyond the lack of a specific arginine residue, disfavor NADPH binding and thus support E2 oxidation by 17βHSD2 in intact cells.
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- 2009
14. Puberty in a case with novel 17-hydroxylase mutation and the putative role of estrogen in development of pubic hair
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Serap Turan, Tulay Guran, Richard J. Auchus, Abdullah Bereket, Teoman Akcay, and Mahboubeh Papari-Zareei
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medicine.medical_specialty ,Adolescent ,medicine.drug_class ,Endocrinology, Diabetes and Metabolism ,Basal (phylogenetics) ,Endocrinology ,Sex hormone-binding globulin ,Internal medicine ,medicine ,Humans ,Point Mutation ,Hydrocortisone ,Menarche ,Breast development ,biology ,Adrenal Hyperplasia, Congenital ,Adrenarche ,Puberty ,Steroid 17-alpha-Hydroxylase ,Estrogens ,General Medicine ,Pubic hair ,medicine.anatomical_structure ,Phenotype ,Estrogen ,CYP17A1 ,biology.protein ,Female ,medicine.drug ,Hair - Abstract
Objective17-Hydroxylase/17,20-lyase deficiency (17OHD) results from mutations in the CYP17A1 gene, leading to failure to synthesize cortisol, adrenal androgens, and gonadal steroids. Adrenarche is a consequence of the increased production of adrenal androgens. Here, we report a case carrying novel R239Q mutation causing complete functional loss of CYP17A1, and thus absence of adrenal and gonadal sex hormone production. The patient has had unexpected pubic hair development and insufficient breast development with estrogen replacement therapy. Possible mechanisms leading to pubic hair development and breast underdevelopment are discussed.Patient and methodsA 15-year-old female born to consanguineous parents presented with the lack of full breast development and irregular menses after the age of 14 years. She had Tanner III breast development on one side, Tanner I on the other side and Tanner I pubic hair and, no axillary hair development. The serum levels of FSH, LH, and progesterone were high and, estradiol was low. The measurement of basal and ACTH-stimulated steroids was consistent with the diagnosis of 17OHD. Genetic analysis revealed novel homozygous mutation R239Q in CYP17A1 gene. Therapy with hydrocortisone was initiated and followed by the addition of conjugated estrogen. Her breast development did not improve considerably, however, pubic hair development started after estrogen treatment in spite of undetectable serum levels of androgens.ConclusionThis case study suggests that estrogen exerts a permissive effect on pubic hair development in girls, even in the presence of very low-circulating androgens, and impaired breast development might be due to estrogen/progesterone imbalance in breast tissue.
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- 2008
15. Cofactors, redox state, and directional preferences of hydroxysteroid dehydrogenases
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Kamalesh K. Sharma, Arhana Chattopadhyay, Anil K. Agarwal, Stefan Andersson, Daniel Sherbet, Mahboubeh Papari-Zareei, Naveed Khan, Richard J. Auchus, Andrew Brandmaier, and Siayareh Rambally
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chemistry.chemical_classification ,Nicotinamide ,Hydroxysteroid Dehydrogenases ,Metabolism ,Biology ,Biochemistry ,Cofactor ,Rats ,chemistry.chemical_compound ,Endocrinology ,Enzyme ,chemistry ,biology.protein ,Animals ,Humans ,NAD+ kinase ,Hydroxysteroid dehydrogenase ,Molecular Biology ,Flux (metabolism) ,Oxidation-Reduction ,NADP - Abstract
The hydroxysteroid dehydrogenases (HSDs) interconvert pairs of weak and potent steroids, thus serving as key enzymes in the regulation of intracellular hormone potency. These enzymes may appear to drive unidirectional steroid flux in intact cells but actually catalyze bi-directional metabolism that achieve pseudo-equilibria with strong directional preferences. Even small shifts in the magnitude of these pseudo-equilibria can profoundly change steroid potency and thus contribute to disease. Consequently, we are studying the structural and biochemical principles that govern these directional preferences and the resilience of these pseudo-equilibria in intact cells. HSD directional preferences in intact cells are governed largely by relative affinities for nicotinamide cofactors [NAD(P)(H)] and existing cofactor gradients. We can attenuate the directional preferences for human 17βHSD type 1 and rat AKR1C9 in intact cells by either diminishing the NADPH/NADP+ gradient or by mutating the arginine residues that form salt bridges with the 2′-phosphate of NADP(H) (R38 and R276, respectively).
- Published
- 2007
16. Abstract 225: Assessing and enriching human tumor cell content in patient-derived cancer xenografts and co-cultures
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Jingsheng Yan, Victor Stastny, John D. Minna, Yuan Zhang, Boning Gao, Brenda C. Timmons, Adi F. Gazdar, Heather L. Davidson, C. Patrick Reynolds, and Mahboubeh Papari-Zareei
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Human tumor ,Cancer Research ,medicine.anatomical_structure ,Oncology ,business.industry ,Cell ,medicine ,Cancer research ,Cancer ,In patient ,business ,Bioinformatics ,medicine.disease - Abstract
Purpose: To develop a robust real-time PCR-based assay with improved accuracy for quantifying the relative fractions of human and mouse DNA in heterogeneous samples such as patient-derived cancer xenografts or cultures. Background: Patient derived mouse xenografts (PDXs), or irradiated mouse fibroblast conditionally reprogrammed cancer cultures (CRCs) provide valuable resources for studying cancers, but as mouse tissue in heterogeneous samples can significantly confound downstream functional and molecular assays (such as next-generation sequencing) it is important to determine the fraction of mouse DNA in mixed samples. Methods: Using an approach different from a previously reported PCR assay (Alcoser SY et al BMC Biotech 2011, 11:124), we established a Taqman real-time PCR assay employing mouse- and human-specific primers that amplified the same chromosomal region of PTGER2 on human chromosome 14q22.1 and mouse chromosome 14 22.68 cM. We also developed a method to enrich human tumor cells from PDXs using enzymatic digestion with Liberase (Roche), followed by differential centrifugation and adherence. Results: Utilizing serial dilutions of heterogeneous DNA mixtures of mouse spleen DNA and normal human white blood cell DNA at varied DNA ratios we demonstrated the assay to be linear (Pearson r = 0.9984, human DNA; r = 0.9998, mouse DNA; p6.4 pg DNA/reaction). The assay was evaluated in 66 samples of human tumors (lung, ovary, neuroblastoma, colon, pancreas, stomach and melanoma) including 25 PDXs before and after tumor cell enrichment, 12 non-enriched PDXs, 2 CRC-derived xenografts and 2 CRCs. Of the PDXs 28 were subcutaneous tumors and 9 were malignant ascites. A wide range of human:mouse DNA ratios were found from zero to 99% among all samples, with two PDXs and 2 CRC-derived xenografts lacking detectable human DNA. The highest human cell fractions were in the malignant ascites samples. Of the enriched samples, 18 of 25 (72%) showed tumor cell enrichment of 2 to 71-fold as compared with the value of matching non-enriched PDX. Conclusions: We developed a robust real-time PCR assay for measuring the relative fraction of species-specific DNA in human:mouse heterogenous samples. The human:mouse DNA ratios in PDX samples varied widely and the human tumor cell populations in PDXs can be effectively enriched in vitro. Assessing human tumor cell content with this assay will facilitate molecular studies with PDXs and mouse-human cocultivated cells. Citation Format: Yu-An Zhang, Victor Stastny, Mahboubeh Papari-Zareei, Heather Davidson, Boning Gao, Brenda Timmons, Jingsheng Yan, C Patrick Reynolds, John D. Minna, Adi F. Gazdar. Assessing and enriching human tumor cell content in patient-derived cancer xenografts and co-cultures. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 225. doi:10.1158/1538-7445.AM2015-225
- Published
- 2015
17. Two novel mutations found in a patient with 17alpha-hydroxylase enzyme deficiency
- Author
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Richard J. Auchus, Berrin Ergun-Longmire, Robert C. Wilson, Susan Tansil, Maria I. New, and Mahboubeh Papari-Zareei
- Subjects
Adult ,Male ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,Context (language use) ,Biology ,medicine.disease_cause ,Biochemistry ,DNA sequencing ,Exon ,Endocrinology ,Internal medicine ,medicine ,Humans ,Congenital adrenal hyperplasia ,Gene ,Genetics ,chemistry.chemical_classification ,Mutation ,Adrenal Hyperplasia, Congenital ,Biochemistry (medical) ,Steroid 17-alpha-Hydroxylase ,medicine.disease ,Molecular biology ,Enzyme assay ,Enzyme ,chemistry ,biology.protein ,Female - Abstract
Context: Congenital adrenal hyperplasia resulting from 17α-hydroxylase deficiency (17OHD) is a rare disorder associated with hypertension. Subject and Methods: We describe a phenotypically and hormonally affected female patient with 17OHD. DNA sequencing of her CYP17 gene revealed a maternal heterozygous mutation in exon 2 (R125Q) and a paternal heterozygous mutation in exon 8 (R416H). These are novel mutations in the CYP17 gene that completely eliminate enzyme activity. Conclusion: Identification of novel mutations in the CYP17 gene is vital in understanding the molecular mechanisms of its deficiency and in providing additional information about the structure and enzymatic functions of P450c17.
- Published
- 2006
18. Abstract 5605: Spectral karyotyping characterization of ovarian adenocarcinomas and corresponding cell lines and xenografts
- Author
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Jayanthi S. Lea, Evelyn Musselwhite, Mahboubeh Papari-Zareei, Adi F. Gazdar, Victor Stastny, Marileila Varella-Garcia, Sakshi Mahale, Tito Woodburn, Patrick Reynolds, and Isabel M. Bernal
- Subjects
Cancer Research ,Pathology ,medicine.medical_specialty ,Breakpoint ,Cancer ,Karyotype ,Ovary ,Biology ,medicine.disease ,Molecular biology ,Serous fluid ,medicine.anatomical_structure ,Oncology ,In vivo ,medicine ,Ovarian cancer ,Ex vivo - Abstract
Ovarian cancer is the 9th leading cancer and the 5th cause of cancer deaths in women in the USA. To understand tumor biology, identify oncogenic driver pathways and verify response to drugs, numerous in vitro (cell lines) and in vivo (xenograft in mouse) models have been developed and extensively used. In this study, we aimed to cytogenetically characterize ovarian carcinomas and their respective in vitro and in vivo models to investigate how representative the models are of their originating source. Ascitic fluid was collected from 10 women with previously untreated high grade serous papillary adenocarcinomas of the ovary that presented with malignant ascites (stage 3). Ascitic fluid was subjected to differential centrifugation and plating to enrich for the tumor cell population (TCP). The TCP cells were cultured and considered permanent cell lines (CLP) when grew continuously without evidence of a non-malignant component and could be recovered from cryopreservation. TCP cells were also inoculated intraperitoneally into athymic nude mice and tumors harvested and cryopreserved (XCP). Eight pairs of TCP and CLP and two triplets of TCP, CLP and XCP specimens were characterized by spectral karyotyping (SKY), for a total of 22 specimens. Highly rearranged karyotypes were detected in all specimens. All matched pairs and triplets were obviously related, sharing most of the structural (SA) and numerical (NA) abnormalities identified. Numerous examples of evolution from simple to complex chromosomal rearrangements and of multiple rearrangements originating from a single breakpoint were detected. For 9 patients, TCP and CLP specimens had the same ploidy level (near-2n in 4, near-3n in 3 and near 4n in 2); for one patient, the ploidy changed from near-2n (TCP) to near-4n (CLP). The 2 xenografts had same ploidy as their matched TCP and CLP specimens. In individual analyses, specific SA accounted for 30% in 6. Specific NA accounted for 30% in 3. In the TCP-CLP analyses, 3 pairs were very similar, differing by 30%. Higher frequency of abnormalities was detected in the TCP specimen for 4 pairs, and in the CLP for 3 pairs. One xenograft was very similar to both TCP and CLP, while the other was closer to the CLP. In conclusion, the ovarian adenocarcinomas were highly rearranged chromosomally and highly heterogeneous. In vitro (CLP) and in vivo (XCP) models maintained a core of the TCP's SA and NA but also displayed unique events, quantitatively and qualitatively variable in distinct tumors, indicating evidence of ex vivo tumor cell selection or progression events. These findings have implications for using established tumor models in research, as well as for potentially identifying the driver rearrangements in specific tumors. Citation Format: Marileila Varella-Garcia, Isabel M. Bernal, Sakshi Mahale, Evelyn M. Musselwhite, Victor Stastny, Mahboubeh Papari-Zareei, Jayanthi Lea, Tito Woodburn, Patrick Reynolds, Adi F. Gazdar. Spectral karyotyping characterization of ovarian adenocarcinomas and corresponding cell lines and xenografts. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5605. doi:10.1158/1538-7445.AM2014-5605
- Published
- 2014
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