32 results on '"Häupl B"'
Search Results
2. Comparative proteomics reveals a diagnostic signature for pulmonary head-and-neck cancer metastasis
- Author
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Bohnenberger, H., Kaderali, L., Ströbel, P., Yepes, D., Pleßmann, U., Dharia, N., Yao, S., Heydt, C., Merkelbach‐Bruse, S., Emmert, A., Hoffmann, J., Bodemeyer, J., Reuter‐Jessen, K., Lois, A., Dröge, L., Baumeister, P., Walz, C., Biggemann, L., Walter, R., Häupl, B., Comoglio, F., Pan, K., Scheich, S., Lenz, C., Küffer, S., Bremmer, F., Kitz, J., Sitte, M., Beißbarth, T., Hinterthaner, M., Sebastian, M., Lotz, J., Schildhaus, H., Wolff, H., Danner, B., Brandts, C., Büttner, R., Canis, M., Stegmaier, K., Serve, H., Urlaub, H., and Oellerich, T.
- Subjects
stomatognathic diseases - Abstract
Patients with head‐and‐neck cancer can develop both lung metastasis and primary lung cancer during the course of their disease. Despite the clinical importance of discrimination, reliable diagnostic biomarkers are still lacking. Here, we have characterised a cohort of squamous cell lung (SQCLC) and head‐and‐neck (HNSCC) carcinomas by quantitative proteomics. In a training cohort, we quantified 4,957 proteins in 44 SQCLC and 30 HNSCC tumours. A total of 518 proteins were found to be differentially expressed between SQCLC and HNSCC, and some of these were identified as genetic dependencies in either of the two tumour types. Using supervised machine learning, we inferred a proteomic signature for the classification of squamous cell carcinomas as either SQCLC or HNSCC, with diagnostic accuracies of 90.5% and 86.8% in cross‐ and independent validations, respectively. Furthermore, application of this signature to a cohort of pulmonary squamous cell carcinomas of unknown origin leads to a significant prognostic separation. This study not only provides a diagnostic proteomic signature for classification of secondary lung tumours in HNSCC patients, but also represents a proteomic resource for HNSCC and SQCLC.
- Published
- 2018
3. Molecular targets of glucocorticoids that elucidate their therapeutic efficacy in aggressive lymphomas.
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Choi J, Ceribelli M, Phelan JD, Häupl B, Huang DW, Wright GW, Hsiao T, Morris V, Ciccarese F, Wang B, Corcoran S, Scheich S, Yu X, Xu W, Yang Y, Zhao H, Zhou J, Zhang G, Muppidi J, Inghirami GG, Oellerich T, Wilson WH, Thomas CJ, and Staudt LM
- Subjects
- Humans, Animals, Signal Transduction drug effects, Receptors, Glucocorticoid metabolism, Mice, Cell Line, Tumor, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse genetics, Lymphoma, Large B-Cell, Diffuse metabolism, Burkitt Lymphoma drug therapy, Burkitt Lymphoma genetics, Burkitt Lymphoma metabolism, Burkitt Lymphoma pathology, Molecular Targeted Therapy methods, Phosphatidylinositol 3-Kinases metabolism, src-Family Kinases metabolism, Gene Expression Regulation, Neoplastic drug effects, Glucocorticoids pharmacology, Receptors, Antigen, B-Cell metabolism
- Abstract
Glucocorticoids have been used for decades to treat lymphomas without an established mechanism of action. Using functional genomic, proteomic, and chemical screens, we discover that glucocorticoids inhibit oncogenic signaling by the B cell receptor (BCR), a recurrent feature of aggressive B cell malignancies, including diffuse large B cell lymphoma and Burkitt lymphoma. Glucocorticoids induce the glucocorticoid receptor (GR) to directly transactivate genes encoding negative regulators of BCR stability (LAPTM5; KLHL14) and the PI3 kinase pathway (INPP5D; DDIT4). GR directly represses transcription of CSK, a kinase that limits the activity of BCR-proximal Src-family kinases. CSK inhibition attenuates the constitutive BCR signaling of lymphomas by hyperactivating Src-family kinases, triggering their ubiquitination and degradation. With the knowledge that glucocorticoids disable oncogenic BCR signaling, they can now be deployed rationally to treat BCR-dependent aggressive lymphomas and used to construct mechanistically sound combination regimens with inhibitors of BTK, PI3 kinase, BCL2, and CSK., Competing Interests: Declaration of interests The authors declare no competing interests., (Published by Elsevier Inc.)
- Published
- 2024
- Full Text
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4. SH2 domain-containing inositol 5-phosphatases support the survival of Burkitt lymphoma cells by promoting energy metabolism.
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Mayr F, Kruse V, Fuhrmann DC, Wolf S, Löber J, Alsouri S, Paglilla N, Lee K, Chapuy B, Brüne B, Zenz T, Häupl B, Oellerich T, and Engelke M
- Subjects
- Humans, Cell Line, Tumor, Apoptosis, src Homology Domains, Signal Transduction, Cell Proliferation, Receptors, Antigen, B-Cell metabolism, Burkitt Lymphoma metabolism, Burkitt Lymphoma genetics, Burkitt Lymphoma pathology, Energy Metabolism, Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases metabolism, Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases genetics, Cell Survival
- Abstract
Burkitt lymphoma cells (BL) exploit antigen-independent tonic signals transduced by the B-cell antigen receptor (BCR) for their survival, but the molecular details of the rewired BL-specific BCR signal network remain unclear. A loss of function screen revealed the SH2 domain-containing 5`-inositol phosphatase 2 (SHIP2) as a potential modulator of BL fitness. We characterized the role of SHIP2 in BL survival in several BL cell models and show that perturbing SHIP2 function renders cells more susceptible to apoptosis, while attenuating proliferation in a BCR-dependent manner. Unexpectedly, SHIP2 deficiency did neither affect PI3K survival signals nor MAPK activity, but attenuated ATP production. We found that an efficient energy metabolism in BL cells requires phosphatidylinositol-3,4-bisphosphate (PI(3,4)P2), which is the enzymatic product of SHIP proteins. Consistently, interference with the function of SHIP1 and SHIP2 augments BL cell susceptibility to PI3K inhibition. Notably, we provide here a molecular basis of how tonic BCR signals are connected to energy supply, which is particularly important for such an aggressively growing neoplasia. These findings may help to improve therapies for the treatment of BL by limiting energy metabolism through the inhibition of SHIP proteins, which renders BL cells more susceptible to the targeting of survival signals.
- Published
- 2024
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5. Response to Bruton's tyrosine kinase inhibitors in aggressive lymphomas linked to chronic selective autophagy.
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Phelan JD, Scheich S, Choi J, Wright GW, Häupl B, Young RM, Rieke SA, Pape M, Ji Y, Urlaub H, Bolomsky A, Doebele C, Zindel A, Wotapek T, Kasbekar M, Collinge B, Huang DW, Coulibaly ZA, Morris VM, Zhuang X, Enssle JC, Yu X, Xu W, Yang Y, Zhao H, Wang Z, Tran AD, Shoemaker CJ, Shevchenko G, Hodson DJ, Shaffer AL 3rd, Staudt LM, and Oellerich T
- Subjects
- Humans, Myeloid Differentiation Factor 88 genetics, Myeloid Differentiation Factor 88 metabolism, Myeloid Differentiation Factor 88 pharmacology, Signal Transduction, Autophagy, Tyrosine Kinase Inhibitors, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse genetics, Lymphoma, Large B-Cell, Diffuse pathology
- Abstract
Diffuse large B cell lymphoma (DLBCL) is an aggressive, profoundly heterogeneous cancer, presenting a challenge for precision medicine. Bruton's tyrosine kinase (BTK) inhibitors block B cell receptor (BCR) signaling and are particularly effective in certain molecular subtypes of DLBCL that rely on chronic active BCR signaling to promote oncogenic NF-κB. The MCD genetic subtype, which often acquires mutations in the BCR subunit, CD79B, and in the innate immune adapter, MYD88
L265P , typically resists chemotherapy but responds exceptionally to BTK inhibitors. However, the underlying mechanisms of response to BTK inhibitors are poorly understood. Herein, we find a non-canonical form of chronic selective autophagy in MCD DLBCL that targets ubiquitinated MYD88L265P for degradation in a TBK1-dependent manner. MCD tumors acquire genetic and epigenetic alterations that attenuate this autophagic tumor suppressive pathway. In contrast, BTK inhibitors promote autophagic degradation of MYD88L265P , thus explaining their exceptional clinical benefit in MCD DLBCL., Competing Interests: Declaration of interests T.O. received research funding from Gilead and Merck KGaA, is a consultant/received honoraria for/from Beigene, Roche, Janssen, Merck KGaA, Gilead, Kronos Bio and Abbvie (all not related to this work). Z.W. is a current employee at GSK. A.L.S III is a current employee at AstraZeneca and has stock options., (Copyright © 2023 Elsevier Inc. All rights reserved.)- Published
- 2024
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6. The MYC-Regulated RNA-Binding Proteins hnRNPC and LARP1 Are Drivers of Multiple Myeloma Cell Growth and Disease Progression and Negatively Predict Patient Survival.
- Author
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Seibert M, Koschade SE, Stolp V, Häupl B, Wempe F, Serve H, Kurrle N, Schnütgen F, and von Metzler I
- Abstract
Multiple myeloma (MM) is a malignant plasma cell disorder in which the MYC oncogene is frequently dysregulated. Due to its central role, MYC has been proposed as a drug target; however, the development of a clinically applicable molecule modulating MYC activity remains an unmet challenge. Consequently, an alternative is the development of therapeutic options targeting proteins located downstream of MYC. Therefore, we aimed to identify undescribed MYC-target proteins in MM cells using Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) and mass spectrometry. We revealed a cluster of proteins associated with the regulation of translation initiation. Herein, the RNA-binding proteins Heterogeneous Nuclear Ribonucleoprotein C (hnRNPC) and La Ribonucleoprotein 1 (LARP1) were predominantly downregulated upon MYC depletion. CRISPR-mediated knockout of either hnRNPC or LARP1 in conjunction with redundant LARP family proteins resulted in a proliferative disadvantage for MM cells. Moreover, high expression levels of these proteins correlate with high MYC expression and with poor survival and disease progression in MM patients. In conclusion, our study provides valuable insights into MYC's role in translation initiation by identifying hnRNPC and LARP1 as proliferation drivers of MM cells and as both predictive factors for survival and disease progression in MM patients., Competing Interests: The authors declare no conflict of interest.
- Published
- 2023
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7. Targeting N-linked Glycosylation for the Therapy of Aggressive Lymphomas.
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Scheich S, Chen J, Liu J, Schnütgen F, Enssle JC, Ceribelli M, Thomas CJ, Choi J, Morris V, Hsiao T, Nguyen H, Wang B, Bolomsky A, Phelan JD, Corcoran S, Urlaub H, Young RM, Häupl B, Wright GW, Huang DW, Ji Y, Yu X, Xu W, Yang Y, Zhao H, Muppidi J, Pan KT, Oellerich T, and Staudt LM
- Subjects
- Humans, Glycosylation, Signal Transduction, Phosphatidylinositol 3-Kinases metabolism, Cell Line, Tumor, NF-kappa B metabolism, Lymphoma, Large B-Cell, Diffuse drug therapy, Lymphoma, Large B-Cell, Diffuse genetics, Lymphoma, Large B-Cell, Diffuse metabolism
- Abstract
Diffuse large B-cell lymphoma (DLBCL) can be subdivided into the activated B-cell (ABC) and germinal center B cell-like (GCB) subtypes. Self-antigen engagement of B-cell receptors (BCR) in ABC tumors induces their clustering, thereby initiating chronic active signaling and activation of NF-κB and PI3 kinase. Constitutive BCR signaling is essential in some GCB tumors but primarily activates PI3 kinase. We devised genome-wide CRISPR-Cas9 screens to identify regulators of IRF4, a direct transcriptional target of NF-κB and an indicator of proximal BCR signaling in ABC DLBCL. Unexpectedly, inactivation of N-linked protein glycosylation by the oligosaccharyltransferase-B (OST-B) complex reduced IRF4 expression. OST-B inhibition of BCR glycosylation reduced BCR clustering and internalization while promoting its association with CD22, which attenuated PI3 kinase and NF-κB activation. By directly interfering with proximal BCR signaling, OST-B inactivation killed models of ABC and GCB DLBCL, supporting the development of selective OST-B inhibitors for the treatment of these aggressive cancers., Significance: DLBCL depends on constitutive BCR activation and signaling. There are currently no therapeutics that target the BCR directly and attenuate its pathologic signaling. Here, we unraveled a therapeutically exploitable, OST-B-dependent glycosylation pathway that drives BCR organization and proximal BCR signaling. This article is highlighted in the In This Issue feature, p. 1749., (©2023 American Association for Cancer Research.)
- Published
- 2023
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8. The RHOA Mutation G17V Does Not Lead to Increased Migration of Human Malignant T Cells but Is Associated with Matrix Remodelling.
- Author
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Merk-Ahmad K, Bein J, Scharf S, Schäfer H, Bexte T, Ullrich E, Loth AG, Flinner N, Senff T, Schneider O, Hansmann ML, Piel M, Häupl B, Oellerich T, Donnadieu E, and Hartmann S
- Abstract
Nodal T-follicular helper cell lymphoma, angioimmunoblastic-type (AITL), is characterized by constitutional symptoms, advanced-stage disease, and generalized lymphadenopathy. A genetic hallmark of this lymphoma is the frequent occurrence of the RHOA mutation G17V in neoplastic cells, which is observed in around 60% of patients. Because RHOA is involved in both T-cell receptor downstream signalling and cell migration, we hypothesized that the characteristic presentation of AITL could be the result of enhanced tumor cell migration. Therefore, this study aimed to elucidate the impact of the RHOA variant G17V on the migration of neoplastic T cells. We transfected the T-cell lymphoma cell lines HH and HuT78 to stably express the RHOA-G17V variant. RHOA-G17V-expressing T cells did not exhibit enhanced motility compared to empty-vector-transfected cells in microchannels, a 3D collagen gel, or primary human lymphatic tissue. Cells of the HH cell line expressing RHOA-G17V had an increased number of cells with cleaved collagen compared with the empty-vector-transfected cells. Therefore, we hypothesized that the early spread of AITL tumor cells may be related to remodelling of the extracellular matrix. Accordingly, we observed a significant negative correlation between the relative area of collagen in histological sections from 18 primary AITL and the allele frequency of the RHOA -G17V mutation. In conclusion, our results suggest that the characteristic presentation of AITL with early, widespread dissemination of lymphoma cells is not the result of an enhanced migration capacity due to the RHOA -G17V mutation; instead, this feature may rather be related to extracellular matrix remodelling.
- Published
- 2023
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9. LYN kinase programs stromal fibroblasts to facilitate leukemic survival via regulation of c-JUN and THBS1.
- Author
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Vom Stein AF, Rebollido-Rios R, Lukas A, Koch M, von Lom A, Reinartz S, Bachurski D, Rose F, Bozek K, Abdallah AT, Kohlhas V, Saggau J, Zölzer R, Zhao Y, Bruns C, Bröckelmann PJ, Lohneis P, Büttner R, Häupl B, Oellerich T, Nguyen PH, and Hallek M
- Subjects
- Humans, Fibroblasts metabolism, Gene Expression Regulation, Leukemic, Leukemia genetics, Signal Transduction, Leukemia, Lymphocytic, Chronic, B-Cell genetics, src-Family Kinases metabolism, Proto-Oncogene Proteins c-jun metabolism, Thrombospondins metabolism
- Abstract
Microenvironmental bystander cells are essential for the progression of chronic lymphocytic leukemia (CLL). We have discovered previously that LYN kinase promotes the formation of a microenvironmental niche for CLL. Here we provide mechanistic evidence that LYN regulates the polarization of stromal fibroblasts to support leukemic progression. LYN is overexpressed in fibroblasts of lymph nodes of CLL patients. LYN-deficient stromal cells reduce CLL growth in vivo. LYN-deficient fibroblasts show markedly reduced leukemia feeding capacity in vitro. Multi-omics profiling reveals that LYN regulates the polarization of fibroblasts towards an inflammatory cancer-associated phenotype through modulation of cytokine secretion and extracellular matrix composition. Mechanistically, LYN deletion reduces inflammatory signaling including reduction of c-JUN expression, which in turn augments the expression of Thrombospondin-1, which binds to CD47 thereby impairing CLL viability. Together, our findings suggest that LYN is essential for rewiring fibroblasts towards a leukemia-supportive phenotype., (© 2023. The Author(s).)
- Published
- 2023
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10. Correction: The non-apoptotic function of Caspase-8 in negatively regulating the CDK9-mediated Ser2 phosphorylation of RNA polymerase II in cervical cancer.
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Mandal R, Raab M, Rödel F, Krämer A, Kostova I, Peña-Llopis S, Medici G, Häupl B, Oellerich T, Gasimli K, Sanhaji M, Becker S, and Strebhardt K
- Published
- 2023
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11. The non-apoptotic function of Caspase-8 in negatively regulating the CDK9-mediated Ser2 phosphorylation of RNA polymerase II in cervical cancer.
- Author
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Mandal R, Raab M, Rödel F, Krämer A, Kostova I, Peña-Llopis S, Häupl B, Oellerich T, Gasimli K, Sanhaji M, Becker S, and Strebhardt K
- Subjects
- Humans, Female, Phosphorylation, Caspase 8 genetics, Caspase 8 metabolism, Cisplatin pharmacology, Protein Kinase Inhibitors, Cyclin-Dependent Kinase 9 genetics, Cyclin-Dependent Kinase 9 metabolism, RNA Polymerase II metabolism, Uterine Cervical Neoplasms drug therapy, Uterine Cervical Neoplasms genetics
- Abstract
Cervical cancer is the fourth most frequently diagnosed and fatal gynecological cancer. 15-61% of all cases metastasize and develop chemoresistance, reducing the 5-year survival of cervical cancer patients to as low as 17%. Therefore, unraveling the mechanisms contributing to metastasis is critical in developing better-targeted therapies against it. Here, we have identified a novel mechanism where nuclear Caspase-8 directly interacts with and inhibits the activity of CDK9, thereby modulating RNAPII-mediated global transcription, including those of cell-migration- and cell-invasion-associated genes. Crucially, low Caspase-8 expression in cervical cancer patients leads to poor prognosis, higher CDK9 phosphorylation at Thr186, and increased RNAPII activity in cervical cancer cell lines and patient biopsies. Caspase-8 knock-out cells were also more resistant to the small-molecule CDK9 inhibitor BAY1251152 in both 2D- and 3D-culture conditions. Combining BAY1251152 with Cisplatin synergistically overcame chemoresistance of Caspase-8-deficient cervical cancer cells. Therefore, Caspase-8 expression could be a marker in chemoresistant cervical tumors, suggesting CDK9 inhibitor treatment for their sensitization to Cisplatin-based chemotherapy., (© 2022. The Author(s).)
- Published
- 2022
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12. Publisher Correction: Oncogenic RAS commandeers amino acid sensing machinery to aberrantly activate mTORC1 in multiple myeloma.
- Author
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Yang Y, Bolomsky A, Oellerich T, Chen P, Ceribelli M, Häupl B, Wright GW, Phelan JD, Huang DW, Lord JW, Van Winkle CK, Yu X, Wisniewski J, Wang JQ, Tosto FA, Beck E, Wilson K, McKnight C, Travers J, Klumpp-Thomas C, Smith GA, Pittaluga S, Maric I, Kazandjian D, Thomas CJ, and Young RM
- Published
- 2022
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13. Oncogenic RAS commandeers amino acid sensing machinery to aberrantly activate mTORC1 in multiple myeloma.
- Author
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Yang Y, Bolomsky A, Oellerich T, Chen P, Ceribelli M, Häupl B, Wright GW, Phelan JD, Huang DW, Lord JW, Van Winkle CK, Yu X, Wisniewski J, Wang JQ, Tosto FA, Beck E, Wilson K, McKnight C, Travers J, Klumpp-Thomas C, Smith GA, Pittaluga S, Maric I, Kazandjian D, Thomas CJ, and Young RM
- Subjects
- Amino Acids metabolism, Humans, Mechanistic Target of Rapamycin Complex 1 genetics, Mechanistic Target of Rapamycin Complex 1 metabolism, Mitogen-Activated Protein Kinase Kinases, Mutation, Protein Isoforms, Genes, ras genetics, Genes, ras physiology, Multiple Myeloma genetics, Multiple Myeloma metabolism, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Oncogenic RAS mutations are common in multiple myeloma (MM), an incurable malignancy of plasma cells. However, the mechanisms of pathogenic RAS signaling in this disease remain enigmatic and difficult to inhibit therapeutically. We employ an unbiased proteogenomic approach to dissect RAS signaling in MM. We discover that mutant isoforms of RAS organize a signaling complex with the amino acid transporter, SLC3A2, and MTOR on endolysosomes, which directly activates mTORC1 by co-opting amino acid sensing pathways. MM tumors with high expression of mTORC1-dependent genes are more aggressive and enriched in RAS mutations, and we detect interactions between RAS and MTOR in MM patient tumors harboring mutant RAS isoforms. Inhibition of RAS-dependent mTORC1 activity synergizes with MEK and ERK inhibitors to quench pathogenic RAS signaling in MM cells. This study redefines the RAS pathway in MM and provides a mechanistic and rational basis to target this mode of RAS signaling., (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)
- Published
- 2022
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14. Early B cell factor 4 modulates FAS-mediated apoptosis and promotes cytotoxic function in human immune cells.
- Author
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Kubo S, Kataria R, Yao Y, Gabrielski JQ, Zheng L, Markowitz TE, Chan W, Song J, Boddapati AK, Saeki K, Häupl B, Park AY, Cheng YH, Cui J, Oellerich T, and Lenardo MJ
- Subjects
- Animals, Chromatin metabolism, Granzymes genetics, Humans, Mice, Perforin metabolism, Apoptosis physiology, CD8-Positive T-Lymphocytes, Cytotoxicity, Immunologic genetics, Fas Ligand Protein metabolism, T-Lymphocytes, Cytotoxic, Transcription Factors genetics, Transcription Factors metabolism
- Abstract
Apoptosis is a genetically regulated program of cell death that plays a key role in immune disease processes. We identified EBF4, a little-studied member of the early B cell factor (EBF) family of transcription factors, in a whole-genome CRISPR screen for regulators of Fas/APO-1/CD95-mediated T cell death. Loss of EBF4 increases the half-life of the c-FLIP protein, and its presence in the Fas signaling complex impairs caspase-8 cleavage and apoptosis. Transcriptome analysis revealed that EBF4 regulates molecules such as TBX21, EOMES, granzyme, and perforin that are important for human natural killer (NK) and CD8
+ T cell functions. Proximity-dependent biotin identification (Bio-ID) mass spectrometry analyses showed EBF4 binding to STAT3, STAT5, and MAP kinase 3 and a strong pathway relationship to interleukin-2 regulated genes, which are known to govern cytotoxicity pathways. Chromatin immunoprecipitation and DNA sequencing analysis defined a canonical EBF4 binding motif, 5'-CCCNNGG/AG-3', closely related to the EBF1 binding site; using a luciferase-based reporter, we found a dose-dependent transcriptional response of this motif to EBF4. We also conducted assay for transposase-accessible chromatin sequencing in EBF4-overexpressing cells and found increased chromatin accessibility upstream of granzyme and perforin and in topologically associated domains in human lymphocytes. Finally, we discovered that the EBF4 has basal expression in human but not mouse NK cells and CD8+ T cells and vanishes following activating stimulation. Together, our data reveal key features of a previously unknown transcriptional regulator of human cytotoxic immune function.- Published
- 2022
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15. Activation of CD44/PAK1/AKT signaling promotes resistance to FGFR1 inhibition in squamous-cell lung cancer.
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Elakad O, Häupl B, Labitzky V, Yao S, Küffer S, von Hammerstein-Equord A, Danner BC, Jücker M, Urlaub H, Lange T, Ströbel P, Oellerich T, and Bohnenberger H
- Abstract
Lung cancer is the leading cause of cancer-related deaths worldwide. Fibroblast growth factor receptor 1 (FGFR1) gene amplification is one of the most prominent and potentially targetable genetic alterations in squamous-cell lung cancer (SQCLC). Highly selective tyrosine kinase inhibitors have been developed to target FGFR1; however, resistance mechanisms originally existing in patients or acquired during treatment have so far led to limited treatment efficiency in clinical trials. In this study we performed a wide-scale phosphoproteomic mass-spectrometry analysis to explore signaling pathways that lead to resistance toward FGFR1 inhibition in lung cancer cells that display (i) intrinsic, (ii) pharmacologically induced and (iii) mutationally induced resistance. Additionally, we correlated AKT activation to CD44 expression in 175 lung cancer patient samples. We identified a CD44/PAK1/AKT signaling axis as a commonly occurring resistance mechanism to FGFR1 inhibition in lung cancer. Co-inhibition of AKT/FGFR1, CD44/FGFR1 or PAK1/FGFR1 sensitized 'intrinsically resistant' and 'induced-resistant' lung-cancer cells synergetically to FGFR1 inhibition. Furthermore, strong CD44 expression was significantly correlated with AKT activation in SQCLC patients. Collectively, our phosphoproteomic analysis of lung-cancer cells resistant to FGFR1 inhibitor provides a large data library of resistance-associated phosphorylation patterns and leads to the proposal of a common resistance pathway comprising CD44, PAK1 and AKT activation. Examination of CD44/PAK1/AKT activation could help to predict response to FGFR1 inhibition. Moreover, combination between AKT and FGFR1 inhibitors may pave the way for an effective therapy of patients with treatment-resistant FGFR1-dependent lung cancer., (© 2022. The Author(s).)
- Published
- 2022
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16. T-cell-derived Hodgkin lymphoma has motility characteristics intermediate between Hodgkin and anaplastic large cell lymphoma.
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Bein J, Flinner N, Häupl B, Mathur A, Schneider O, Abu-Ayyad M, Hansmann ML, Piel M, Oellerich T, and Hartmann S
- Subjects
- Humans, Proteomics, T-Lymphocytes metabolism, Hodgkin Disease genetics, Hodgkin Disease metabolism, Hodgkin Disease pathology, Lymphoma, Large-Cell, Anaplastic metabolism, Lymphoma, Large-Cell, Anaplastic pathology
- Abstract
Classic Hodgkin lymphoma (cHL) is usually characterized by a low tumour cell content, derived from crippled germinal centre B cells. Rare cases have been described in which the tumour cells show clonal T-cell receptor rearrangements. From a clinicopathological perspective, it is unclear if these cases should be classified as cHL or anaplastic large T-cell lymphoma (ALCL). Since we recently observed differences in the motility of ALCL and cHL tumour cells, here, we aimed to obtain a better understanding of T-cell-derived cHL by investigating their global proteomic profiles and their motility. In a proteomics analysis, when only motility-associated proteins were regarded, T-cell-derived cHL cell lines showed the highest similarity to ALK
- ALCL cell lines. In contrast, T-cell-derived cHL cell lines presented a very low overall motility, similar to that observed in conventional cHL. Whereas all ALCL cell lines, as well as T-cell-derived cHL, predominantly presented an amoeboid migration pattern with uropod at the rear, conventional cHL never presented with uropods. The migration of ALCL cell lines was strongly impaired upon application of different inhibitors. This effect was less pronounced in cHL cell lines and almost invisible in T-cell-derived cHL. In summary, our cell line-derived data suggest that based on proteomics and migration behaviour, T-cell-derived cHL is a neoplasm that shares features with both cHL and ALCL and is not an ALCL with low tumour cell content. Complementary clinical studies on this lymphoma are warranted., (© 2022 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)- Published
- 2022
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17. The proteogenomic subtypes of acute myeloid leukemia.
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Jayavelu AK, Wolf S, Buettner F, Alexe G, Häupl B, Comoglio F, Schneider C, Doebele C, Fuhrmann DC, Wagner S, Donato E, Andresen C, Wilke AC, Zindel A, Jahn D, Splettstoesser B, Plessmann U, Münch S, Abou-El-Ardat K, Makowka P, Acker F, Enssle JC, Cremer A, Schnütgen F, Kurrle N, Chapuy B, Löber J, Hartmann S, Wild PJ, Wittig I, Hübschmann D, Kaderali L, Cox J, Brüne B, Röllig C, Thiede C, Steffen B, Bornhäuser M, Trumpp A, Urlaub H, Stegmaier K, Serve H, Mann M, and Oellerich T
- Subjects
- Humans, Proteomics, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute pathology, Proteogenomics
- Abstract
Acute myeloid leukemia (AML) is an aggressive blood cancer with a poor prognosis. We report a comprehensive proteogenomic analysis of bone marrow biopsies from 252 uniformly treated AML patients to elucidate the molecular pathophysiology of AML in order to inform future diagnostic and therapeutic approaches. In addition to in-depth quantitative proteomics, our analysis includes cytogenetic profiling and DNA/RNA sequencing. We identify five proteomic AML subtypes, each reflecting specific biological features spanning genomic boundaries. Two of these proteomic subtypes correlate with patient outcome, but none is exclusively associated with specific genomic aberrations. Remarkably, one subtype (Mito-AML), which is captured only in the proteome, is characterized by high expression of mitochondrial proteins and confers poor outcome, with reduced remission rate and shorter overall survival on treatment with intensive induction chemotherapy. Functional analyses reveal that Mito-AML is metabolically wired toward stronger complex I-dependent respiration and is more responsive to treatment with the BCL2 inhibitor venetoclax., Competing Interests: Declaration of interests T.O. received research funding from Gilead (related to this work) and Merck KGaA (not related to this work). T.O. is a consultant for Roche and Merck KGaA (both not related to this work). K.S. receives grant funding as part of the DFCI/Novartis Drug Discovery Program, consults for and has stock options in Auron Therapeutics, and has consulted for Kronos Bio and AstraZeneca on topics not directly related to this manuscript. F.C. is a co-founder of enGene Statistics GmbH. The Max Planck institute and the Goethe University Frankfurt are filing a patent application, on which T.O., A.K.J., S.Wolf, F.B., H.S., M.M., and H.U. are listed as inventors., (Copyright © 2022 Elsevier Inc. All rights reserved.)
- Published
- 2022
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18. Inflammatory fibroblasts mediate resistance to neoadjuvant therapy in rectal cancer.
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Nicolas AM, Pesic M, Engel E, Ziegler PK, Diefenhardt M, Kennel KB, Buettner F, Conche C, Petrocelli V, Elwakeel E, Weigert A, Zinoveva A, Fleischmann M, Häupl B, Karakütük C, Bohnenberger H, Mosa MH, Kaderali L, Gaedcke J, Ghadimi M, Rödel F, Arkan MC, Oellerich T, Rödel C, Fokas E, and Greten FR
- Subjects
- Animals, Biomarkers, Cancer-Associated Fibroblasts pathology, Cell Line, Tumor, Cellular Senescence drug effects, Cellular Senescence genetics, Cytokines genetics, Cytokines metabolism, DNA Damage, Disease Models, Animal, Disease Susceptibility, Gene Expression Profiling, Heterografts, High-Throughput Nucleotide Sequencing, Humans, Immunohistochemistry, Kaplan-Meier Estimate, Mice, Neoadjuvant Therapy, Prognosis, Rectal Neoplasms drug therapy, Rectal Neoplasms etiology, Rectal Neoplasms pathology, Signal Transduction, Cancer-Associated Fibroblasts metabolism, Drug Resistance, Neoplasm, Rectal Neoplasms metabolism, Tumor Microenvironment genetics
- Abstract
Standard cancer therapy targets tumor cells without considering possible damage on the tumor microenvironment that could impair therapy response. In rectal cancer patients we find that inflammatory cancer-associated fibroblasts (iCAFs) are associated with poor chemoradiotherapy response. Employing a murine rectal cancer model or patient-derived tumor organoids and primary stroma cells, we show that, upon irradiation, interleukin-1α (IL-1α) not only polarizes cancer-associated fibroblasts toward the inflammatory phenotype but also triggers oxidative DNA damage, thereby predisposing iCAFs to p53-mediated therapy-induced senescence, which in turn results in chemoradiotherapy resistance and disease progression. Consistently, IL-1 inhibition, prevention of iCAFs senescence, or senolytic therapy sensitizes mice to irradiation, while lower IL-1 receptor antagonist serum levels in rectal patients correlate with poor prognosis. Collectively, we unravel a critical role for iCAFs in rectal cancer therapy resistance and identify IL-1 signaling as an attractive target for stroma-repolarization and prevention of cancer-associated fibroblasts senescence., Competing Interests: Declaration of interests The authors declare no conflict of interests. The authors have filed a patent application related to the study., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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19. SHMT2 inhibition disrupts the TCF3 transcriptional survival program in Burkitt lymphoma.
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Wilke AC, Doebele C, Zindel A, Lee KS, Rieke SA, Ceribelli M, Comoglio F, Phelan JD, Wang JQ, Pikman Y, Jahn D, Häupl B, Schneider C, Scheich S, Tosto FA, Bohnenberger H, Stauder P, Schnütgen F, Slabicki M, Coulibaly ZA, Wolf S, Bojarczuk K, Chapuy B, Brandts CH, Stroebel P, Lewis CA, Engelke M, Xu X, Kim H, Dang TH, Schmitz R, Hodson DJ, Stegmaier K, Urlaub H, Serve H, Schmitt CA, Kreuz F, Knittel G, Rabinowitz JD, Reinhardt HC, Vander Heiden MG, Thomas C, Staudt LM, Zenz T, and Oellerich T
- Subjects
- Animals, Burkitt Lymphoma genetics, Cell Line, Tumor, Cell Survival drug effects, Drug Discovery, Formates metabolism, Gene Expression Regulation, Neoplastic drug effects, Gene Knockdown Techniques, Glycine metabolism, Glycine Hydroxymethyltransferase genetics, Humans, Mice, Molecular Targeted Therapy, Proteolysis drug effects, Basic Helix-Loop-Helix Transcription Factors metabolism, Burkitt Lymphoma drug therapy, Burkitt Lymphoma metabolism, Glycine Hydroxymethyltransferase antagonists & inhibitors, Glycine Hydroxymethyltransferase metabolism
- Abstract
Burkitt lymphoma (BL) is an aggressive lymphoma type that is currently treated by intensive chemoimmunotherapy. Despite the favorable clinical outcome for most patients with BL, chemotherapy-related toxicity and disease relapse remain major clinical challenges, emphasizing the need for innovative therapies. Using genome-scale CRISPR-Cas9 screens, we identified B-cell receptor (BCR) signaling, specific transcriptional regulators, and one-carbon metabolism as vulnerabilities in BL. We focused on serine hydroxymethyltransferase 2 (SHMT2), a key enzyme in one-carbon metabolism. Inhibition of SHMT2 by either knockdown or pharmacological compounds induced anti-BL effects in vitro and in vivo. Mechanistically, SHMT2 inhibition led to a significant reduction of intracellular glycine and formate levels, which inhibited the mTOR pathway and thereby triggered autophagic degradation of the oncogenic transcription factor TCF3. Consequently, this led to a collapse of tonic BCR signaling, which is controlled by TCF3 and is essential for BL cell survival. In terms of clinical translation, we also identified drugs such as methotrexate that synergized with SHMT inhibitors. Overall, our study has uncovered the dependency landscape in BL, identified and validated SHMT2 as a drug target, and revealed a mechanistic link between SHMT2 and the transcriptional master regulator TCF3, opening up new perspectives for innovative therapies., (© 2022 by The American Society of Hematology.)
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- 2022
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20. Identification of the Cysteine Protease Legumain as a Potential Chronic Hypoxia-Specific Multiple Myeloma Target Gene.
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Clees AS, Stolp V, Häupl B, Fuhrmann DC, Wempe F, Seibert M, Weber S, Banning A, Tikkanen R, Williams R, Brüne B, Serve H, Schnütgen F, von Metzler I, and Kurrle N
- Subjects
- Basic Helix-Loop-Helix Transcription Factors metabolism, CRISPR-Cas Systems genetics, Cell Line, Tumor, Cell Proliferation genetics, Gene Expression Regulation, Neoplastic, Hexokinase metabolism, Humans, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, Lactate Dehydrogenase 5 metabolism, Proteome metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, Signal Transduction genetics, Up-Regulation genetics, Cysteine Endopeptidases genetics, Molecular Targeted Therapy, Multiple Myeloma enzymology, Multiple Myeloma genetics, Tumor Hypoxia genetics
- Abstract
Multiple myeloma (MM) is the second most common hematologic malignancy, which is characterized by clonal proliferation of neoplastic plasma cells in the bone marrow. This microenvironment is characterized by low oxygen levels (1-6% O
2 ), known as hypoxia. For MM cells, hypoxia is a physiologic feature that has been described to promote an aggressive phenotype and to confer drug resistance. However, studies on hypoxia are scarce and show little conformity. Here, we analyzed the mRNA expression of previously determined hypoxia markers to define the temporal adaptation of MM cells to chronic hypoxia. Subsequent analyses of the global proteome in MM cells and the stromal cell line HS-5 revealed hypoxia-dependent regulation of proteins, which directly or indirectly upregulate glycolysis. In addition, chronic hypoxia led to MM-specific regulation of nine distinct proteins. One of these proteins is the cysteine protease legumain (LGMN), the depletion of which led to a significant growth disadvantage of MM cell lines that is enhanced under hypoxia. Thus, herein, we report a methodologic strategy to examine MM cells under physiologic hypoxic conditions in vitro and to decipher and study previously masked hypoxia-specific therapeutic targets such as the cysteine protease LGMN.- Published
- 2022
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21. Overcoming Acquired Epigenetic Resistance to BTK Inhibitors.
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Shaffer AL 3rd, Phelan JD, Wang JQ, Huang D, Wright GW, Kasbekar M, Choi J, Young RM, Webster DE, Yang Y, Zhao H, Yu X, Xu W, Roulland S, Ceribelli M, Zhang X, Wilson KM, Chen L, McKnight C, Klumpp-Thomas C, Thomas CJ, Häupl B, Oellerich T, Rae Z, Kelly MC, Ahn IE, Sun C, Gaglione EM, Wilson WH, Wiestner A, and Staudt LM
- Subjects
- Agammaglobulinaemia Tyrosine Kinase genetics, Drug Resistance, Neoplasm genetics, Epigenesis, Genetic, Humans, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy, Protein Kinase Inhibitors pharmacology
- Abstract
The use of Bruton tyrosine kinase (BTK) inhibitors to block B-cell receptor (BCR)-dependent NF-κB activation in lymphoid malignancies has been a major clinical advance, yet acquired therapeutic resistance is a recurring problem. We modeled the development of resistance to the BTK inhibitor ibrutinib in the activated B-cell (ABC) subtype of diffuse large B-cell lymphoma, which relies on chronic active BCR signaling for survival. The primary mode of resistance was epigenetic, driven in part by the transcription factor TCF4. The resultant phenotypic shift altered BCR signaling such that the GTPase RAC2 substituted for BTK in the activation of phospholipase Cγ2, thereby sustaining NF-κB activity. The interaction of RAC2 with phospholipase Cγ2 was also increased in chronic lymphocytic leukemia cells from patients with persistent or progressive disease on BTK inhibitor treatment. We identified clinically available drugs that can treat epigenetic ibrutinib resistance, suggesting combination therapeutic strategies., Significance: In diffuse large B-cell lymphoma, we show that primary resistance to BTK inhibitors is due to epigenetic rather than genetic changes that circumvent the BTK blockade. We also observed this resistance mechanism in chronic lymphocytic leukemia, suggesting that epigenetic alterations may contribute more to BTK inhibitor resistance than currently thought. See related commentary by Pasqualucci, p. 555 . This article is highlighted in the In This Issue feature, p. 549 ., (©2021 American Association for Cancer Research.)
- Published
- 2021
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22. PRMT6 activates cyclin D1 expression in conjunction with the transcription factor LEF1.
- Author
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Schneider L, Herkt S, Wang L, Feld C, Wesely J, Kuvardina ON, Meyer A, Oellerich T, Häupl B, Seifried E, Bonig H, and Lausen J
- Abstract
The establishment of cell type specific gene expression by transcription factors and their epigenetic cofactors is central for cell fate decisions. Protein arginine methyltransferase 6 (PRMT6) is an epigenetic regulator of gene expression mainly through methylating arginines at histone H3. This way it influences cellular differentiation and proliferation. PRMT6 lacks DNA-binding capability but is recruited by transcription factors to regulate gene expression. However, currently only a limited number of transcription factors have been identified, which facilitate recruitment of PRMT6 to key cell cycle related target genes. Here, we show that LEF1 contributes to the recruitment of PRMT6 to the central cell cycle regulator CCND1 (Cyclin D1). We identified LEF1 as an interaction partner of PRMT6. Knockdown of LEF1 or PRMT6 reduces CCND1 expression. This is in line with our observation that knockdown of PRMT6 increases the number of cells in G1 phase of the cell cycle and decreases proliferation. These results improve the understanding of PRMT6 activity in cell cycle regulation. We expect that these insights will foster the rational development and usage of specific PRMT6 inhibitors for cancer therapy.
- Published
- 2021
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23. The transcription factor TAL1 and miR-17-92 create a regulatory loop in hematopoiesis.
- Author
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Meyer A, Herkt S, Kunze-Schumacher H, Kohrs N, Ringleb J, Schneider L, Kuvardina ON, Oellerich T, Häupl B, Krueger A, Seifried E, Bonig H, and Lausen J
- Subjects
- Feedback, Physiological, Gene Expression Regulation, HEK293 Cells, Hematopoiesis, Humans, Jurkat Cells, K562 Cells, Protein Stability, RNA, Long Noncoding, T-Cell Acute Lymphocytic Leukemia Protein 1 chemistry, Transcription Factor 3 metabolism, Transcriptional Activation, Erythroid Cells cytology, MicroRNAs genetics, T-Cell Acute Lymphocytic Leukemia Protein 1 genetics, T-Cell Acute Lymphocytic Leukemia Protein 1 metabolism
- Abstract
A network of gene regulatory factors such as transcription factors and microRNAs establish and maintain gene expression patterns during hematopoiesis. In this network, transcription factors regulate each other and are involved in regulatory loops with microRNAs. The microRNA cluster miR-17-92 is located within the MIR17HG gene and encodes six mature microRNAs. It is important for hematopoietic differentiation and plays a central role in malignant disease. However, the transcription factors downstream of miR-17-92 are largely elusive and the transcriptional regulation of miR-17-92 is not fully understood. Here we show that miR-17-92 forms a regulatory loop with the transcription factor TAL1. The miR-17-92 cluster inhibits expression of TAL1 and indirectly leads to decreased stability of the TAL1 transcriptional complex. We found that TAL1 and its heterodimerization partner E47 regulate miR-17-92 transcriptionally. Furthermore, miR-17-92 negatively influences erythroid differentiation, a process that depends on gene activation by the TAL1 complex. Our data give example of how transcription factor activity is fine-tuned during normal hematopoiesis. We postulate that disturbance of the regulatory loop between TAL1 and the miR-17-92 cluster could be an important step in cancer development and progression.
- Published
- 2020
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24. Metabolic Plasticity Is an Essential Requirement of Acquired Tyrosine Kinase Inhibitor Resistance in Chronic Myeloid Leukemia.
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Mostazo MGC, Kurrle N, Casado M, Fuhrmann D, Alshamleh I, Häupl B, Martín-Sanz P, Brüne B, Serve H, Schwalbe H, Schnütgen F, Marin S, and Cascante M
- Abstract
Tyrosine kinase inhibitors (TKIs) are currently the standard chemotherapeutic agents for the treatment of chronic myeloid leukemia (CML). However, due to TKI resistance acquisition in CML patients, identification of new vulnerabilities is urgently required for a sustained response to therapy. In this study, we have investigated metabolic reprogramming induced by TKIs independent of BCR-ABL1 alterations. Proteomics and metabolomics profiling of imatinib-resistant CML cells (ImaR) was performed. KU812 ImaR cells enhanced pentose phosphate pathway, glycogen synthesis, serine-glycine-one-carbon metabolism, proline synthesis and mitochondrial respiration compared with their respective syngeneic parental counterparts. Moreover, the fact that only 36% of the main carbon sources were utilized for mitochondrial respiration pointed to glycerol-phosphate shuttle as mainly contributors to mitochondrial respiration. In conclusion, CML cells that acquire TKIs resistance present a severe metabolic reprogramming associated with an increase in metabolic plasticity needed to overcome TKI-induced cell death. Moreover, this study unveils that KU812 Parental and ImaR cells viability can be targeted with metabolic inhibitors paving the way to propose novel and promising therapeutic opportunities to overcome TKI resistance in CML.
- Published
- 2020
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25. AKT-dependent NOTCH3 activation drives tumor progression in a model of mesenchymal colorectal cancer.
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Varga J, Nicolas A, Petrocelli V, Pesic M, Mahmoud A, Michels BE, Etlioglu E, Yepes D, Häupl B, Ziegler PK, Bankov K, Wild PJ, Wanninger S, Medyouf H, Farin HF, Tejpar S, Oellerich T, Ruland J, Siebel CW, and Greten FR
- Subjects
- Animals, Colorectal Neoplasms pathology, Disease Models, Animal, Disease Progression, Gene Expression Regulation, Neoplastic, Humans, Mice, Mice, Inbred C57BL, Neoplasm Transplantation, Signal Transduction, Transcriptome, Up-Regulation, Colorectal Neoplasms metabolism, Proto-Oncogene Proteins c-akt metabolism, Receptor, Notch3 metabolism
- Abstract
Recently, a transcriptome-based consensus molecular subtype (CMS) classification of colorectal cancer (CRC) has been established, which may ultimately help to individualize CRC therapy. However, the lack of animal models that faithfully recapitulate the different molecular subtypes impedes adequate preclinical testing of stratified therapeutic concepts. Here, we demonstrate that constitutive AKT activation in intestinal epithelial cells markedly enhances tumor invasion and metastasis in Trp53ΔIEC mice (Trp53ΔIECAktE17K) upon challenge with the carcinogen azoxymethane. Gene-expression profiling indicates that Trp53ΔIECAktE17K tumors resemble the human mesenchymal colorectal cancer subtype (CMS4), which is characterized by the poorest survival rate among the four CMSs. Trp53ΔIECAktE17K tumor cells are characterized by Notch3 up-regulation, and treatment of Trp53ΔIECAktE17K mice with a NOTCH3-inhibiting antibody reduces invasion and metastasis. In CRC patients, NOTCH3 expression correlates positively with tumor grading and the presence of lymph node as well as distant metastases and is specifically up-regulated in CMS4 tumors. Therefore, we suggest NOTCH3 as a putative target for advanced CMS4 CRC patients., Competing Interests: Disclosures: T. Oellerich reported grants from Merck KGaA, grants from Gilead, personal fees from Merck KGaA, and personal fees from Gilead outside the submitted work. C. Siebel reported personal fees from Genentech during the conduct of the study, and personal fees from Genentech outside the submitted work. In addition, C. Siebel had a patent to Methods of treating cancer using Notch1 and Notch3 antagonists issued. No other disclosures were reported., (© 2020 Varga et al.)
- Published
- 2020
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26. Regulation of B cell receptor-dependent NF-κB signaling by the tumor suppressor KLHL14.
- Author
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Choi J, Phelan JD, Wright GW, Häupl B, Huang DW, Shaffer AL 3rd, Young RM, Wang Z, Zhao H, Yu X, Oellerich T, and Staudt LM
- Subjects
- Adenine analogs & derivatives, CD79 Antigens genetics, Carrier Proteins metabolism, Cell Line, Tumor, Drug Resistance, Neoplasm genetics, Endoplasmic Reticulum metabolism, HEK293 Cells, Humans, Intracellular Signaling Peptides and Proteins, Lymphoma, Large B-Cell, Diffuse pathology, Mutagenesis, Site-Directed, Myeloid Differentiation Factor 88 metabolism, NF-kappa B metabolism, Piperidines, Proteolysis, Pyrazoles pharmacology, Pyrazoles therapeutic use, Pyrimidines pharmacology, Pyrimidines therapeutic use, Signal Transduction drug effects, Signal Transduction genetics, Carrier Proteins genetics, Genes, Tumor Suppressor, Lymphoma, Large B-Cell, Diffuse genetics, Receptors, Antigen, B-Cell metabolism, Ubiquitin-Protein Ligase Complexes metabolism
- Abstract
The KLHL14 gene acquires frequent inactivating mutations in mature B cell malignancies, especially in the MYD88
L265P , CD79B mutant (MCD) genetic subtype of diffuse large B cell lymphoma (DLBCL), which relies on B cell receptor (BCR) signaling for survival. However, the pathogenic role of KLHL14 in DLBCL and its molecular function are largely unknown. Here, we report that KLHL14 is in close proximity to the BCR in the endoplasmic reticulum of MCD cell line models and promotes the turnover of immature glycoforms of BCR subunits, reducing total cellular BCR levels. Loss of KLHL14 confers relative resistance to the Bruton tyrosine kinase (BTK) inhibitor ibrutinib and promotes assembly of the MYD88-TLR9-BCR (My-T-BCR) supercomplex, which initiates prosurvival NF-κB activation. Consequently, KLHL14 inactivation allows MCD cells to maintain NF-κB signaling in the presence of ibrutinib. These findings reinforce the central role of My-T-BCR-dependent NF-κB signaling in MCD DLBCL and suggest that the genetic status of KLHL14 should be considered in clinical trials testing inhibitors of BTK and BCR signaling mediators in DLBCL., Competing Interests: The authors declare no competing interest.- Published
- 2020
- Full Text
- View/download PDF
27. Resistance Mechanisms to SYK Inhibition in Acute Myeloid Leukemia.
- Author
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Cremer A, Ellegast JM, Alexe G, Frank ES, Ross L, Chu SH, Pikman Y, Robichaud A, Goodale A, Häupl B, Mohr S, Rao AV, Walker AR, Blachly JS, Piccioni F, Armstrong SA, Byrd JC, Oellerich T, and Stegmaier K
- Subjects
- Animals, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Benzamides pharmacology, Benzamides therapeutic use, Cell Line, Tumor, Clinical Trials, Phase I as Topic, Clinical Trials, Phase II as Topic, Diphenylamine analogs & derivatives, Diphenylamine pharmacology, Diphenylamine therapeutic use, Drug Resistance, Neoplasm drug effects, Drug Synergism, Female, Gene Expression Regulation, Leukemic drug effects, Humans, Indazoles pharmacology, Indazoles therapeutic use, Leukemia, Myeloid, Acute genetics, MAP Kinase Signaling System drug effects, MAP Kinase Signaling System genetics, Mice, Mitogen-Activated Protein Kinase 1 genetics, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinase Kinases metabolism, Mutagenesis, Site-Directed, Mutation, Open Reading Frames genetics, Primary Cell Culture, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Protein Tyrosine Phosphatase, Non-Receptor Type 11 genetics, Protein Tyrosine Phosphatase, Non-Receptor Type 11 metabolism, Pyrazines pharmacology, Pyrazines therapeutic use, Syk Kinase metabolism, Xenograft Model Antitumor Assays, Antineoplastic Combined Chemotherapy Protocols pharmacology, Drug Resistance, Neoplasm genetics, Leukemia, Myeloid, Acute drug therapy, Mitogen-Activated Protein Kinase Kinases antagonists & inhibitors, Syk Kinase antagonists & inhibitors
- Abstract
Spleen tyrosine kinase (SYK) is a nonmutated therapeutic target in acute myeloid leukemia (AML). Attempts to exploit SYK therapeutically in AML have shown promising results in combination with chemotherapy, likely reflecting induced mechanisms of resistance to single-agent treatment in vivo . We conducted a genome-scale open reading frame (ORF) resistance screen and identified activation of the RAS-MAPK-ERK pathway as one major mechanism of resistance to SYK inhibitors. This finding was validated in AML cell lines with innate and acquired resistance to SYK inhibitors. Furthermore, patients with AML with select mutations activating these pathways displayed early resistance to SYK inhibition. To circumvent SYK inhibitor therapy resistance in AML, we demonstrate that a MEK and SYK inhibitor combination is synergistic in vitro and in vivo . Our data provide justification for use of ORF screening to identify resistance mechanisms to kinase inhibitor therapy in AML lacking distinct mutations and to direct novel combination-based strategies to abrogate these. SIGNIFICANCE: The integration of functional genomic screening with the study of mechanisms of intrinsic and acquired resistance in model systems and human patients identified resistance to SYK inhibitors through MAPK signaling in AML. The dual targeting of SYK and the MAPK pathway offers a combinatorial strategy to overcome this resistance. This article is highlighted in the In This Issue feature, p. 161 ., (©2019 American Association for Cancer Research.)
- Published
- 2020
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28. Integrated analysis of relapsed B-cell precursor Acute Lymphoblastic Leukemia identifies subtype-specific cytokine and metabolic signatures.
- Author
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Schroeder MP, Bastian L, Eckert C, Gökbuget N, James AR, Tanchez JO, Schlee C, Isaakidis K, Häupl B, Baum K, Migueles Lozano OA, Kouidri K, Pan KT, Urlaub H, Schwartz S, Burmeister T, von Stackelberg A, Hoelzer D, Pfeiffer H, Rieger MA, Göllner S, Oellerich T, Horstman M, Schrappe M, Wolf J, Kirschner-Schwabe R, Brüggemann M, Müller-Tidow C, Serve H, Neumann M, and Baldus CD
- Subjects
- Adolescent, Adult, Child, Female, Genomics, Humans, Male, Metabolic Networks and Pathways, Proteomics, Cytokines genetics, Cytokines metabolism, Gene Expression Regulation, Leukemic, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma classification, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma genetics, Precursor B-Cell Lymphoblastic Leukemia-Lymphoma metabolism
- Abstract
Recent efforts reclassified B-Cell Precursor Acute Lymphoblastic Leukemia (BCP-ALL) into more refined subtypes. Nevertheless, outcomes of relapsed BCP-ALL remain unsatisfactory, particularly in adult patients where the molecular basis of relapse is still poorly understood. To elucidate the evolution of relapse in BCP-ALL, we established a comprehensive multi-omics dataset including DNA-sequencing, RNA-sequencing, DNA methylation array and proteome MASS-spec data from matched diagnosis and relapse samples of BCP-ALL patients (n = 50) including the subtypes DUX4, Ph-like and two aneuploid subtypes. Relapse-specific alterations were enriched for chromatin modifiers, nucleotide and steroid metabolism including the novel candidates FPGS, AGBL and ZNF483. The proteome expression analysis unraveled deregulation of metabolic pathways at relapse including the key proteins G6PD, TKT, GPI and PGD. Moreover, we identified a novel relapse-specific gene signature specific for DUX4 BCP-ALL patients highlighting chemotaxis and cytokine environment as a possible driver event at relapse. This study presents novel insights at distinct molecular levels of relapsed BCP-ALL based on a comprehensive multi-omics integrated data set including a valuable proteomics data set. The relapse specific aberrations reveal metabolic signatures on genomic and proteomic levels in BCP-ALL relapse. Furthermore, the chemokine expression signature in DUX4 relapse underscores the distinct status of DUX4-fusion BCP-ALL.
- Published
- 2019
- Full Text
- View/download PDF
29. Phosphoproteomic Analysis of Signaling Pathways in Lymphomas.
- Author
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Häupl B, Urlaub H, and Oellerich T
- Subjects
- Humans, Phosphopeptides analysis, Phosphopeptides metabolism, Phosphoproteins analysis, Chromatography, High Pressure Liquid methods, Lymphoma metabolism, Phosphoproteins metabolism, Proteomics methods, Signal Transduction, Tandem Mass Spectrometry methods
- Abstract
Cell fate decisions are controlled by complex signal transduction processes that transmit information via posttranslational protein modifications such as phosphorylation. In lymphoma, as in other cancer types, these signaling networks are often dysregulated and thus contribute to malignant transformation and tumor maintenance. For example, B-cell antigen receptor signals are rewired in certain lymphoma types, such as diffuse large B-cell lymphomas, to promote cell growth and survival of the malignant cell clones. Hence, global elucidation of such intricate signaling networks is important for an improved understanding of the biology of these tumors and the identification of target proteins for therapeutic purposes.We describe here a mass spectrometry-based phosphoproteomic approach for characterization of intracellular signaling events and their dynamics. This integrated phosphoproteomic technology combines phosphopeptide enrichment and fractionation with liquid-chromatography-coupled mass spectrometry for the site-specific mapping and quantification of thousands of phosphorylation events in a given cell type. Such global signaling analyses provide valuable insights into oncogenic signaling networks and can inform drug development efforts.
- Published
- 2019
- Full Text
- View/download PDF
30. Comparative proteomics reveals a diagnostic signature for pulmonary head-and-neck cancer metastasis.
- Author
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Bohnenberger H, Kaderali L, Ströbel P, Yepes D, Plessmann U, Dharia NV, Yao S, Heydt C, Merkelbach-Bruse S, Emmert A, Hoffmann J, Bodemeyer J, Reuter-Jessen K, Lois AM, Dröge LH, Baumeister P, Walz C, Biggemann L, Walter R, Häupl B, Comoglio F, Pan KT, Scheich S, Lenz C, Küffer S, Bremmer F, Kitz J, Sitte M, Beißbarth T, Hinterthaner M, Sebastian M, Lotz J, Schildhaus HU, Wolff H, Danner BC, Brandts C, Büttner R, Canis M, Stegmaier K, Serve H, Urlaub H, and Oellerich T
- Subjects
- Carcinoma, Squamous Cell pathology, Diagnostic Tests, Routine methods, Head and Neck Neoplasms pathology, Humans, Lung Neoplasms pathology, Machine Learning, Sensitivity and Specificity, Carcinoma, Squamous Cell diagnosis, Carcinoma, Squamous Cell secondary, Head and Neck Neoplasms diagnosis, Head and Neck Neoplasms secondary, Lung Neoplasms diagnosis, Proteome analysis, Proteomics methods
- Abstract
Patients with head-and-neck cancer can develop both lung metastasis and primary lung cancer during the course of their disease. Despite the clinical importance of discrimination, reliable diagnostic biomarkers are still lacking. Here, we have characterised a cohort of squamous cell lung (SQCLC) and head-and-neck (HNSCC) carcinomas by quantitative proteomics. In a training cohort, we quantified 4,957 proteins in 44 SQCLC and 30 HNSCC tumours. A total of 518 proteins were found to be differentially expressed between SQCLC and HNSCC, and some of these were identified as genetic dependencies in either of the two tumour types. Using supervised machine learning, we inferred a proteomic signature for the classification of squamous cell carcinomas as either SQCLC or HNSCC, with diagnostic accuracies of 90.5% and 86.8% in cross- and independent validations, respectively. Furthermore, application of this signature to a cohort of pulmonary squamous cell carcinomas of unknown origin leads to a significant prognostic separation. This study not only provides a diagnostic proteomic signature for classification of secondary lung tumours in HNSCC patients, but also represents a proteomic resource for HNSCC and SQCLC., (© 2018 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2018
- Full Text
- View/download PDF
31. Combining affinity enrichment, cross-linking with photo amino acids, and mass spectrometry for probing protein kinase D2 interactions.
- Author
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Häupl B, Ihling CH, and Sinz A
- Abstract
We present a novel approach that relies on the affinity capture of protein interaction partners from a complex mixture, followed by their covalent fixation via UV-induced activation of incorporated diazirine photoreactive amino acids (photo-methionine and photo-leucine). The captured protein complexes are enzymatically digested and interacting proteins are identified and quantified by label-free LC/MS analysis. Using HeLa cell lysates with photo-methionine and photo-leucine-labeled proteins, we were able to capture and preserve protein interactions that are otherwise elusive in conventional pull-down experiments. Our approach is exemplified for mapping the protein interaction network of protein kinase D2, but has the potential to be applied to any protein system. Data are available via ProteomeXchange with identifiers PXD005346 (photo amino acid incorporation) and PXD005349 (enrichment experiments)., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
- Published
- 2017
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- View/download PDF
32. Protein Interaction Network of Human Protein Kinase D2 Revealed by Chemical Cross-Linking/Mass Spectrometry.
- Author
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Häupl B, Ihling CH, and Sinz A
- Subjects
- Carrier Proteins analysis, Carrier Proteins metabolism, Cells, Cultured, Cross-Linking Reagents, Cytosol chemistry, Golgi Apparatus chemistry, Humans, Mass Spectrometry methods, Protein Kinase D2, Subcellular Fractions chemistry, Actin-Related Protein 2 metabolism, Actin-Related Protein 3 metabolism, Protein Interaction Maps, Protein Kinases metabolism
- Abstract
We investigated the interaction network of human PKD2 in the cytosol and in Golgi-enriched subcellular protein fractions by an affinity enrichment strategy combined with chemical cross-linking/mass spectrometry (MS). Analysis of the subproteomes revealed the presence of distinct proteins in the cytosolic and Golgi fractions. The covalent fixation of transient or weak interactors by chemical cross-linking allowed capturing interaction partners that might otherwise disappear during conventional pull-down experiments. In total, 31 interaction partners were identified for PKD2, including glycogen synthase kinase-3 beta (GSK3B), 14-3-3 protein gamma (YWHAG), and the alpha isoform of 55 kDa regulatory subunit B of protein phosphatase 2A (PPP2R2A). Remarkably, the entire seven-subunit Arp2/3 complex (ARPC1B, ARPC2, ARPC3, ARPC4, ARPC5, ACTR3, ACTR2) as well as ARPC1A and ARPC5L, which are putative substitutes of ARPC1B and ARPC5, were identified. We provide evidence of a direct protein-protein interaction between PKD2 and Arp2/3. Our findings will pave the way for further structural and functional studies of PKD2 complexes, especially the PKD2/Arp2/3 interaction, to elucidate the role of PKD2 for transport processes at the trans-Golgi network. Data are available via ProteomeXchange with identifiers PXD003909 (enrichment from cytosolic fractions), PXD003913 (enrichment from Golgi fractions), and PXD003917 (subcellular fractionation).
- Published
- 2016
- Full Text
- View/download PDF
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