48 results on '"Drake, Justin M."'
Search Results
2. CRABP1-complexes in exosome secretion
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Nhieu, Jennifer, Wei, Chin-Wen, Ludwig, Megan, Drake, Justin M., and Wei, Li-Na
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- 2024
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3. ALAN is a computational approach that interprets genomic findings in the context of tumor ecosystems
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Bergom, Hannah E., Shabaneh, Ashraf, Day, Abderrahman, Ali, Atef, Boytim, Ella, Tape, Sydney, Lozada, John R., Shi, Xiaolei, Kerkvliet, Carlos Perez, McSweeney, Sean, Pitzen, Samuel P., Ludwig, Megan, Antonarakis, Emmanuel S., Drake, Justin M., Dehm, Scott M., Ryan, Charles J., Wang, Jinhua, and Hwang, Justin
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- 2023
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4. Targeting RET Kinase in Neuroendocrine Prostate Cancer
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VanDeusen, Halena R, Ramroop, Johnny R, Morel, Katherine L, Bae, Song Yi, Sheahan, Anjali V, Sychev, Zoi, Lau, Nathan A, Cheng, Larry C, Tan, Victor M, Li, Zhen, Petersen, Ashley, Lee, John K, Park, Jung Wook, Yang, Rendong, Hwang, Justin H, Coleman, Ilsa, Witte, Owen N, Morrissey, Colm, Corey, Eva, Nelson, Peter S, Ellis, Leigh, and Drake, Justin M
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Biotechnology ,Aging ,Prostate Cancer ,Cancer ,Clinical Research ,Urologic Diseases ,Development of treatments and therapeutic interventions ,5.1 Pharmaceuticals ,Animals ,Carcinoma ,Neuroendocrine ,Cell Line ,Tumor ,Gene Expression Regulation ,Neoplastic ,Gene Knockdown Techniques ,Heterocyclic Compounds ,4 or More Rings ,Humans ,Male ,Mice ,PC-3 Cells ,Phosphorylation ,Prostatic Neoplasms ,Proteomics ,Proto-Oncogene Proteins c-ret ,Receptors ,Androgen ,Up-Regulation ,Xenograft Model Antitumor Assays ,Oncology and Carcinogenesis ,Developmental Biology ,Oncology & Carcinogenesis - Abstract
The increased treatment of metastatic castration-resistant prostate cancer (mCRPC) with second-generation antiandrogen therapies (ADT) has coincided with a greater incidence of lethal, aggressive variant prostate cancer (AVPC) tumors that have lost dependence on androgen receptor (AR) signaling. These AR-independent tumors may also transdifferentiate to express neuroendocrine lineage markers and are termed neuroendocrine prostate cancer (NEPC). Recent evidence suggests kinase signaling may be an important driver of NEPC. To identify targetable kinases in NEPC, we performed global phosphoproteomics comparing several AR-independent to AR-dependent prostate cancer cell lines and identified multiple altered signaling pathways, including enrichment of RET kinase activity in the AR-independent cell lines. Clinical NEPC patient samples and NEPC patient-derived xenografts displayed upregulated RET transcript and RET pathway activity. Genetic knockdown or pharmacologic inhibition of RET kinase in multiple mouse and human models of NEPC dramatically reduced tumor growth and decreased cell viability. Our results suggest that targeting RET in NEPC tumors with high RET expression could be an effective treatment option. Currently, there are limited treatment options for patients with aggressive neuroendocrine prostate cancer and none are curative. IMPLICATIONS: Identification of aberrantly expressed RET kinase as a driver of tumor growth in multiple models of NEPC provides a significant rationale for testing the clinical application of RET inhibitors in patients with AVPC.
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- 2020
5. MEK-ERK signaling is a therapeutic target in metastatic castration resistant prostate cancer
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Nickols, Nicholas G, Nazarian, Ramin, Zhao, Shuang G, Tan, Victor, Uzunangelov, Vladislav, Xia, Zheng, Baertsch, Robert, Neeman, Elad, Gao, Allen C, Thomas, George V, Howard, Lauren, De Hoedt, Amanda M, Stuart, Josh, Goldstein, Theodore, Chi, Kim, Gleave, Martin E, Graff, Julie N, Beer, Tomasz M, Drake, Justin M, Evans, Christopher P, Aggarwal, Rahul, Foye, Adam, Feng, Felix Y, Small, Eric J, Aronson, William J, Freedland, Stephen J, Witte, Owen N, Huang, Jiaoti, Alumkal, Joshi J, Reiter, Robert E, and Rettig, Matthew B
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Biomedical and Clinical Sciences ,Clinical Sciences ,Oncology and Carcinogenesis ,Prostate Cancer ,Cancer ,Urologic Diseases ,Genetics ,Development of treatments and therapeutic interventions ,5.1 Pharmaceuticals ,Aged ,Antineoplastic Agents ,Biopsy ,Disease-Free Survival ,Gene Amplification ,Gene Expression Regulation ,Neoplastic ,Humans ,MAP Kinase Signaling System ,Male ,Middle Aged ,Mitogen-Activated Protein Kinase 3 ,Molecular Targeted Therapy ,Phosphorylation ,Prospective Studies ,Prostate ,Prostatic Neoplasms ,Castration-Resistant ,Protein Kinase Inhibitors ,Pyridones ,Pyrimidinones ,RNA-Seq ,Urology & Nephrology ,Clinical sciences ,Oncology and carcinogenesis - Abstract
BackgroundMetastatic castration resistant prostate cancer (mCRPC) is incurable and progression after drugs that target the androgen receptor-signaling axis is inevitable. Thus, there is an urgent need to develop more effective treatments beyond hormonal manipulation. We sought to identify activated kinases in mCRPC as therapeutic targets for existing, approved agents, with the goal of identifying candidate drugs for rapid translation into proof of concept Phase II trials in mCRPC.MethodsTo identify evidence of activation of druggable kinases in these patients, we compared mRNA expression from metastatic biopsies of patients with mCRPC (n = 101) to mRNA expression in localized prostate from TCGA and used this analysis to infer differential kinase activity. In addition, we assessed the differential phosphorylation levels for key MAPK pathway kinases between mCRPC and localized prostate cancers.ResultsTranscriptomic profiling of 101 patients with mCRPC as compared to patients with localized prostate cancer identified evidence of hyperactive ERK1, and whole genome sequencing revealed frequent amplifications of members of the MAPK pathway in 32% of this cohort. Next, we confirmed elevated levels of phosphorylated ERK1/2 in castration resistant prostate cancer as compared to untreated primary prostate cancer. We observed that the presence of detectable phosphorylated ERK1/2 in the primary tumor is associated with biochemical failure after radical prostatectomy independent of clinicopathologic features. ERK1 is the immediate downstream target of MEK1/2, which is druggable with trametinib, an approved therapeutic for melanoma. Trametinib elicited a profound biochemical and clinical response in a patient who had failed multiple prior treatments for mCRPC.ConclusionsWe conclude that pharmacologic targeting of the MEK/ERK pathway may be a viable treatment strategy for patients with refractory metastatic prostate cancer. An ongoing Phase II trial tests this hypothesis.
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- 2019
6. DNA-Dependent Protein Kinase Drives Prostate Cancer Progression through Transcriptional Regulation of the Wnt Signaling Pathway
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Kothari, Vishal, Goodwin, Jonathan F, Zhao, Shuang G, Drake, Justin M, Yin, Yi, Chang, S Laura, Evans, Joseph R, Wilder-Romans, Kari, Gabbara, Kristina, Dylgjeri, Emanuela, Chou, Jonathan, Sun, Grace, Tomlins, Scott A, Mehra, Rohit, Hege, Kristen, Filvaroff, Ellen H, Schaeffer, Edward M, Karnes, R Jeffrey, Quigley, David A, Rathkopf, Dana E, He, Housheng H, Speers, Corey, Spratt, Daniel E, Gilbert, Luke A, Ashworth, Alan, Chinnaiyan, Arul M, Raj, Ganesh V, Knudsen, Karen E, and Feng, Felix Y
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Prostate Cancer ,Urologic Diseases ,Genetics ,Aging ,Cancer ,2.1 Biological and endogenous factors ,Aetiology ,Animals ,Biomarkers ,Tumor ,Cell Line ,Tumor ,Cell Movement ,DNA-Activated Protein Kinase ,Disease Models ,Animal ,Disease Progression ,Gene Expression Profiling ,Gene Expression Regulation ,Neoplastic ,Gene Knockdown Techniques ,Heterografts ,Humans ,Male ,Mice ,Neoplasm Metastasis ,Phenotype ,Prostatic Neoplasms ,Protein Binding ,RNA ,Small Interfering ,Transcription ,Genetic ,Wnt Signaling Pathway ,Oncology and Carcinogenesis ,Oncology & Carcinogenesis - Abstract
PURPOSE:Protein kinases are known to play a prominent role in oncogenic progression across multiple cancer subtypes, yet their role in prostate cancer progression remains underexplored. The purpose of this study was to identify kinases that drive prostate cancer progression.Experimental Design: To discover kinases that drive prostate cancer progression, we investigated the association between gene expression of all known kinases and long-term clinical outcomes in tumor samples from 545 patients with high-risk disease. We evaluated the impact of genetic and pharmacologic inhibition of the most significant kinase associated with metastatic progression in vitro and in vivo. RESULTS:DNA-dependent protein kinase (DNAPK) was identified as the most significant kinase associated with metastatic progression in high-risk prostate cancer. Inhibition of DNAPK suppressed the growth of both AR-dependent and AR-independent prostate cancer cells. Gene set enrichment analysis nominated Wnt as the top pathway associated with DNAPK. We found that DNAPK interacts with the Wnt transcription factor LEF1 and is critical for LEF1-mediated transcription. CONCLUSIONS:Our data show that DNAPK drives prostate cancer progression through transcriptional regulation of Wnt signaling and is an attractive therapeutic target in aggressive prostate cancer.
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- 2019
7. Integrative molecular analyses define correlates of high B7-H3 expression in metastatic castrate-resistant prostate cancer
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Shi, Xiaolei, Day, Abderrahman, Bergom, Hannah E., Tape, Sydney, Baca, Sylvan C., Sychev, Zoi E., Larson, Gabrianne, Bozicevich, Asha, Drake, Justin M., Zorko, Nicholas, Wang, Jinhua, Ryan, Charles J., Antonarakis, Emmanuel S., and Hwang, Justin
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- 2022
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8. Cabozantinib and dasatinib synergize to induce tumor regression in non-clear cell renal cell carcinoma
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Lue, Hui-wen, Derrick, Daniel S., Rao, Soumya, Van Gaest, Ahna, Cheng, Larry, Podolak, Jennifer, Lawson, Samantha, Xue, Changhui, Garg, Devin, White, Ralph, III, Ryan, Christopher W., Drake, Justin M., Ritz, Anna, Heiser, Laura M., and Thomas, George V.
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- 2021
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9. Metabolic reprogramming ensures cancer cell survival despite oncogenic signaling blockade
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Lue, Hui-wen, Podolak, Jennifer, Kolahi, Kevin, Cheng, Larry, Rao, Soumya, Garg, Devin, Xue, Chang-Hui, Rantala, Juha K, Tyner, Jeffrey W, Thornburg, Kent L, Martinez-Acevedo, Ann, Liu, Jen-Jane, Amling, Christopher L, Truillet, Charles, Louie, Sharon M, Anderson, Kimberly E, Evans, Michael J, O'Donnell, Valerie B, Nomura, Daniel K, Drake, Justin M, Ritz, Anna, and Thomas, George V
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Cancer ,5.1 Pharmaceuticals ,Development of treatments and therapeutic interventions ,Animals ,Antineoplastic Agents ,Apoptosis ,Autophagy ,Benzamides ,Cell Line ,Tumor ,Cell Respiration ,Cell Survival ,Heterocyclic Compounds ,3-Ring ,Humans ,Lipid Droplets ,Mice ,Mitochondria ,Neoplasms ,Phosphatidylinositol 3-Kinases ,Phosphoinositide-3 Kinase Inhibitors ,Phospholipase A2 Inhibitors ,Phospholipids ,Protein Kinase Inhibitors ,Proto-Oncogene Proteins c-akt ,Pyrimidines ,Signal Transduction ,Tumor Cells ,Cultured ,autophagy ,cancer ,metabolism ,phospholipid ,resistance ,signaling ,Biological Sciences ,Medical and Health Sciences ,Psychology and Cognitive Sciences ,Developmental Biology - Abstract
There is limited knowledge about the metabolic reprogramming induced by cancer therapies and how this contributes to therapeutic resistance. Here we show that although inhibition of PI3K-AKT-mTOR signaling markedly decreased glycolysis and restrained tumor growth, these signaling and metabolic restrictions triggered autophagy, which supplied the metabolites required for the maintenance of mitochondrial respiration and redox homeostasis. Specifically, we found that survival of cancer cells was critically dependent on phospholipase A2 (PLA2) to mobilize lysophospholipids and free fatty acids to sustain fatty acid oxidation and oxidative phosphorylation. Consistent with this, we observed significantly increased lipid droplets, with subsequent mobilization to mitochondria. These changes were abrogated in cells deficient for the essential autophagy gene ATG5 Accordingly, inhibition of PLA2 significantly decreased lipid droplets, decreased oxidative phosphorylation, and increased apoptosis. Together, these results describe how treatment-induced autophagy provides nutrients for cancer cell survival and identifies novel cotreatment strategies to override this survival advantage.
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- 2017
10. Activation of Notch1 synergizes with multiple pathways in promoting castration-resistant prostate cancer
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Stoyanova, Tanya, Riedinger, Mireille, Lin, Shu, Faltermeier, Claire M, Smith, Bryan A, Zhang, Kelvin X, Going, Catherine C, Goldstein, Andrew S, Lee, John K, Drake, Justin M, Rice, Meghan A, Hsu, En-Chi, Nowroozizadeh, Behdokht, Castor, Brandon, Orellana, Sandra Y, Blum, Steven M, Cheng, Donghui, Pienta, Kenneth J, Reiter, Robert E, Pitteri, Sharon J, Huang, Jiaoti, and Witte, Owen N
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Biomedical and Clinical Sciences ,Clinical Sciences ,Oncology and Carcinogenesis ,Urologic Diseases ,Prostate Cancer ,Cancer ,Aging ,Amyloid Precursor Protein Secretases ,Animals ,Biomarkers ,Cell Line ,Tumor ,Cell Nucleus ,Disease Models ,Animal ,Disease Progression ,Epithelial-Mesenchymal Transition ,Gene Expression ,Gene Expression Profiling ,Heterografts ,Humans ,Immunohistochemistry ,Male ,Mice ,Mitogen-Activated Protein Kinases ,Neoplasm Grading ,Neoplasm Metastasis ,Phenotype ,Prostatic Neoplasms ,Castration-Resistant ,Proto-Oncogene Proteins c-akt ,Proto-Oncogene Proteins c-myc ,Receptor ,Notch1 ,Signal Transduction ,Tumor Burden ,raf Kinases ,ras Proteins ,prostate ,cancer ,Notch1 - Abstract
Metastatic castration-resistant prostate cancer (CRPC) is the primary cause of prostate cancer-specific mortality. Defining new mechanisms that can predict recurrence and drive lethal CRPC is critical. Here, we demonstrate that localized high-risk prostate cancer and metastatic CRPC, but not benign prostate tissues or low/intermediate-risk prostate cancer, express high levels of nuclear Notch homolog 1, translocation-associated (Notch1) receptor intracellular domain. Chronic activation of Notch1 synergizes with multiple oncogenic pathways altered in early disease to promote the development of prostate adenocarcinoma. These tumors display features of epithelial-to-mesenchymal transition, a cellular state associated with increased tumor aggressiveness. Consistent with its activation in clinical CRPC, tumors driven by Notch1 intracellular domain in combination with multiple pathways altered in prostate cancer are metastatic and resistant to androgen deprivation. Our study provides functional evidence that the Notch1 signaling axis synergizes with alternative pathways in promoting metastatic CRPC and may represent a new therapeutic target for advanced prostate cancer.
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- 2016
11. Phosphoproteome Integration Reveals Patient-Specific Networks in Prostate Cancer.
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Drake, Justin M, Paull, Evan O, Graham, Nicholas A, Lee, John K, Smith, Bryan A, Titz, Bjoern, Stoyanova, Tanya, Faltermeier, Claire M, Uzunangelov, Vladislav, Carlin, Daniel E, Fleming, Daniel Teo, Wong, Christopher K, Newton, Yulia, Sudha, Sud, Vashisht, Ajay A, Huang, Jiaoti, Wohlschlegel, James A, Graeber, Thomas G, Witte, Owen N, and Stuart, Joshua M
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Humans ,Phosphoproteins ,Proteome ,Signal Transduction ,Algorithms ,Male ,Transcriptome ,Prostatic Neoplasms ,Castration-Resistant ,Precision Medicine ,Prostatic Neoplasms ,Castration-Resistant ,Developmental Biology ,Biological Sciences ,Medical and Health Sciences - Abstract
We used clinical tissue from lethal metastatic castration-resistant prostate cancer (CRPC) patients obtained at rapid autopsy to evaluate diverse genomic, transcriptomic, and phosphoproteomic datasets for pathway analysis. Using Tied Diffusion through Interacting Events (TieDIE), we integrated differentially expressed master transcriptional regulators, functionally mutated genes, and differentially activated kinases in CRPC tissues to synthesize a robust signaling network consisting of druggable kinase pathways. Using MSigDB hallmark gene sets, six major signaling pathways with phosphorylation of several key residues were significantly enriched in CRPC tumors after incorporation of phosphoproteomic data. Individual autopsy profiles developed using these hallmarks revealed clinically relevant pathway information potentially suitable for patient stratification and targeted therapies in late stage prostate cancer. Here, we describe phosphorylation-based cancer hallmarks using integrated personalized signatures (pCHIPS) that shed light on the diversity of activated signaling pathways in metastatic CRPC while providing an integrative, pathway-based reference for drug prioritization in individual patients.
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- 2016
12. Functional screen identifies kinases driving prostate cancer visceral and bone metastasis
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Faltermeier, Claire M, Drake, Justin M, Clark, Peter M, Smith, Bryan A, Zong, Yang, Volpe, Carmen, Mathis, Colleen, Morrissey, Colm, Castor, Brandon, Huang, Jiaoti, and Witte, Owen N
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Biomedical and Clinical Sciences ,Clinical Sciences ,Oncology and Carcinogenesis ,Aging ,Prostate Cancer ,Urologic Diseases ,Cancer ,Aetiology ,2.1 Biological and endogenous factors ,Animals ,Bone Neoplasms ,Bone and Bones ,Cell Line ,Tumor ,Gene Expression Profiling ,Humans ,Lentivirus ,Lung ,Male ,Mice ,Mice ,SCID ,Neoplasm Proteins ,Phosphoproteins ,Prostatic Neoplasms ,Protein Kinases ,Proteomics ,Viscera ,src-Family Kinases ,kinases ,metastasis ,prostate cancer ,bone metastasis - Abstract
Mutationally activated kinases play an important role in the progression and metastasis of many cancers. Despite numerous oncogenic alterations implicated in metastatic prostate cancer, mutations of kinases are rare. Several lines of evidence suggest that nonmutated kinases and their pathways are involved in prostate cancer progression, but few kinases have been mechanistically linked to metastasis. Using a mass spectrometry-based phosphoproteomics dataset in concert with gene expression analysis, we selected over 100 kinases potentially implicated in human metastatic prostate cancer for functional evaluation. A primary in vivo screen based on overexpression of candidate kinases in murine prostate cells identified 20 wild-type kinases that promote metastasis. We queried these 20 kinases in a secondary in vivo screen using human prostate cells. Strikingly, all three RAF family members, MERTK, and NTRK2 drove the formation of bone and visceral metastasis confirmed by positron-emission tomography combined with computed tomography imaging and histology. Immunohistochemistry of tissue microarrays indicated that these kinases are highly expressed in human metastatic castration-resistant prostate cancer tissues. Our functional studies reveal the strong capability of select wild-type protein kinases to drive critical steps of the metastatic cascade, and implicate these kinases in possible therapeutic intervention.
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- 2016
13. DNA-PKcs-Mediated Transcriptional Regulation Drives Prostate Cancer Progression and Metastasis
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Goodwin, Jonathan F, Kothari, Vishal, Drake, Justin M, Zhao, Shuang, Dylgjeri, Emanuela, Dean, Jeffry L, Schiewer, Matthew J, McNair, Christopher, Jones, Jennifer K, Aytes, Alvaro, Magee, Michael S, Snook, Adam E, Zhu, Ziqi, Den, Robert B, Birbe, Ruth C, Gomella, Leonard G, Graham, Nicholas A, Vashisht, Ajay A, Wohlschlegel, James A, Graeber, Thomas G, Karnes, R Jeffrey, Takhar, Mandeep, Davicioni, Elai, Tomlins, Scott A, Abate-Shen, Cory, Sharifi, Nima, Witte, Owen N, Feng, Felix Y, and Knudsen, Karen E
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Biomedical and Clinical Sciences ,Clinical Sciences ,Oncology and Carcinogenesis ,Genetics ,Cancer ,2.1 Biological and endogenous factors ,Animals ,Cell Line ,Tumor ,DNA-Activated Protein Kinase ,DNA-Binding Proteins ,Gene Expression Regulation ,Neoplastic ,Gene Regulatory Networks ,Humans ,Male ,Mice ,Molecular Sequence Data ,Neoplasm Invasiveness ,Neoplasm Transplantation ,Nuclear Proteins ,Prostatic Neoplasms ,Receptors ,Androgen ,Neurosciences ,Oncology & Carcinogenesis ,Biochemistry and cell biology ,Oncology and carcinogenesis - Abstract
Emerging evidence demonstrates that the DNA repair kinase DNA-PKcs exerts divergent roles in transcriptional regulation of unsolved consequence. Here, in vitro and in vivo interrogation demonstrate that DNA-PKcs functions as a selective modulator of transcriptional networks that induce cell migration, invasion, and metastasis. Accordingly, suppression of DNA-PKcs inhibits tumor metastases. Clinical assessment revealed that DNA-PKcs is significantly elevated in advanced disease and independently predicts for metastases, recurrence, and reduced overall survival. Further investigation demonstrated that DNA-PKcs in advanced tumors is highly activated, independent of DNA damage indicators. Combined, these findings reveal unexpected DNA-PKcs functions, identify DNA-PKcs as a potent driver of tumor progression and metastases, and nominate DNA-PKcs as a therapeutic target for advanced malignancies.
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- 2015
14. Clinical Targeting of Mutated and Wild-Type Protein Tyrosine Kinases in Cancer
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Drake, Justin M, Lee, John K, and Witte, Owen N
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Genetics ,Urologic Diseases ,Cancer ,2.1 Biological and endogenous factors ,Aetiology ,Agammaglobulinaemia Tyrosine Kinase ,Animals ,Antineoplastic Agents ,Fusion Proteins ,bcr-abl ,Humans ,Leukemia ,Lymphoma ,Molecular Targeted Therapy ,Mutation ,Protein Kinase Inhibitors ,Protein-Tyrosine Kinases ,Biological Sciences ,Medical and Health Sciences ,Developmental Biology - Abstract
Clinical therapies for cancer have evolved from toxic, nontargeted agents to manageable, highly targeted therapies. Protein tyrosine kinases are a family of signaling molecules implicated in nearly every cancer type and are the foundation for the development of modern targeted agents. Recent genomic analyses have identified activating mutations, translocations, and amplifications of tyrosine kinases. Selective targeting of these genetically altered tyrosine kinases has resulted in significant clinical advances, including increased patient survival. This indicates that altered protein tyrosine kinases are the main drivers of many different cancers. However, lost during analyses of genetic lesions are the contributions of activated, wild-type kinases on tumor-dependent pathways. New approaches in phosphoproteomic technologies have identified several wild-type tyrosine kinase activation states, suggesting that non-genetically altered kinases can be essential "nodes" for signal transduction. Here, we summarize the evidence supporting the common mechanisms of protein tyrosine kinase activation in cancer and provide a personal perspective on the kinases BCR-ABL and BTK, as well as nonmutated kinase targets in prostate cancer, through our work. We outline the mechanisms of tyrosine kinase activation in the absence of direct mutation and discuss whether non-genetically altered tyrosine kinases or their associated downstream signaling pathways can be effectively targeted.
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- 2014
15. Dissemination of Circulating Tumor Cells in Breast and Prostate Cancer: Implications for Early Detection.
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Bae, Song Yi, Kamalanathan, Kaylee Judith, Galeano-Garces, Catalina, Konety, Badrinath R, Antonarakis, Emmanuel S, Parthasarathy, Jayant, Hong, Jiarong, and Drake, Justin M
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CIRCULATING tumor DNA ,PROSTATE cancer ,BLOOD vessels - Abstract
Burgeoning evidence suggests that circulating tumor cells (CTCs) may disseminate into blood vessels at an early stage, seeding metastases in various cancers such as breast and prostate cancer. Simultaneously, the early-stage CTCs that settle in metastatic sites [termed disseminated tumor cells (DTCs)] can enter dormancy, marking a potential source of late recurrence and therapy resistance. Thus, the presence of these early CTCs poses risks to patients but also holds potential benefits for early detection and treatment and opportunities for possibly curative interventions. This review delves into the role of early DTCs in driving latent metastasis within breast and prostate cancer, emphasizing the importance of early CTC detection in these diseases. We further explore the correlation between early CTC detection and poor prognoses, which contribute significantly to increased cancer mortality. Consequently, the detection of CTCs at an early stage emerges as a critical imperative for enhancing clinical diagnostics and allowing for early interventions. [ABSTRACT FROM AUTHOR]
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- 2024
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16. Prostate cancer originating in basal cells progresses to adenocarcinoma propagated by luminal-like cells
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Stoyanova, Tanya, Cooper, Aaron R, Drake, Justin M, Liu, Xian, Armstrong, Andrew J, Pienta, Kenneth J, Zhang, Hong, Kohn, Donald B, Huang, Jiaoti, Witte, Owen N, and Goldstein, Andrew S
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Genetics ,Stem Cell Research ,Aging ,Prostate Cancer ,Cancer ,Stem Cell Research - Nonembryonic - Human ,Rare Diseases ,Urologic Diseases ,Aetiology ,2.1 Biological and endogenous factors ,Adenocarcinoma ,Blotting ,Western ,Cell Transformation ,Neoplastic ,Eukaryotic Initiation Factor-4E ,Flow Cytometry ,Gene Expression Regulation ,Neoplastic ,Humans ,Immunohistochemistry ,Male ,Neoplasms ,Basal Cell ,Phenotype ,Prostatic Neoplasms ,Signal Transduction ,beta Catenin - Abstract
The relationship between the cells that initiate cancer and the cancer stem-like cells that propagate tumors has been poorly defined. In a human prostate tissue transformation model, basal cells expressing the oncogenes Myc and myristoylated AKT can initiate heterogeneous tumors. Tumors contain features of acinar-type adenocarcinoma with elevated eIF4E-driven protein translation and squamous cell carcinoma marked by activated beta-catenin. Lentiviral integration site analysis revealed that alternative histological phenotypes can be clonally derived from a common cell of origin. In advanced disease, adenocarcinoma can be propagated by self-renewing tumor cells with an androgen receptor-low immature luminal phenotype in the absence of basal-like cells. These data indicate that advanced prostate adenocarcinoma initiated in basal cells can be maintained by luminal-like tumor-propagating cells. Determining the cells that maintain human prostate adenocarcinoma and the signaling pathways characterizing these tumor-propagating cells is critical for developing effective therapeutic strategies against this population.
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- 2013
17. Metastatic castration-resistant prostate cancer reveals intrapatient similarity and interpatient heterogeneity of therapeutic kinase targets
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Drake, Justin M, Graham, Nicholas A, Lee, John K, Stoyanova, Tanya, Faltermeier, Claire M, Sud, Sudha, Titz, Björn, Huang, Jiaoti, Pienta, Kenneth J, Graeber, Thomas G, and Witte, Owen N
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Cancer ,Urologic Diseases ,Prostate Cancer ,2.1 Biological and endogenous factors ,Aetiology ,Blotting ,Western ,Cell Line ,Tumor ,Drug Discovery ,Enzyme Activation ,ErbB Receptors ,Humans ,Male ,Mass Spectrometry ,Neoplasm Metastasis ,Phosphoproteins ,Phosphorylation ,Phosphotyrosine ,Precision Medicine ,Principal Component Analysis ,Prostatic Neoplasms ,Castration-Resistant ,Protein-Tyrosine Kinases ,metastasis ,resistance ,personalized medicine ,combination therapy ,phosphotyrosine - Abstract
In prostate cancer, multiple metastases from the same patient share similar copy number, mutational status, erythroblast transformation specific (ETS) rearrangements, and methylation patterns supporting their clonal origins. Whether actionable targets such as tyrosine kinases are also similarly expressed and activated in anatomically distinct metastatic lesions of the same patient is not known. We evaluated active kinases using phosphotyrosine peptide enrichment and quantitative mass spectrometry to identify druggable targets in metastatic castration-resistant prostate cancer obtained at rapid autopsy. We identified distinct phosphopeptide patterns in metastatic tissues compared with treatment-naive primary prostate tissue and prostate cancer cell line-derived xenografts. Evaluation of metastatic castration-resistant prostate cancer samples for tyrosine phosphorylation and upstream kinase targets revealed SRC, epidermal growth factor receptor (EGFR), rearranged during transfection (RET), anaplastic lymphoma kinase (ALK), and MAPK1/3 and other activities while exhibiting intrapatient similarity and interpatient heterogeneity. Phosphoproteomic analyses and identification of kinase activation states in metastatic castration-resistant prostate cancer patients have allowed for the prioritization of kinases for further clinical evaluation.
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- 2013
18. Integrating phosphoproteomics into the clinical management of prostate cancer
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Cheng, Larry C., Tan, Victor M., Ganesan, Shridar, and Drake, Justin M.
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- 2017
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19. PIP5K1α inhibition as a therapeutic strategy for prostate cancer
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Drake, Justin M. and Huang, Jiaoti
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- 2014
20. Saracatinib synergizes with enzalutamide to downregulate AR activity in CRPC.
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WhiteIII, Ralph E., Bannister, Maxwell, Day, Abderrahman, Bergom, Hannah E., Tan, Victor M., Hwang, Justin, Hai Dang Nguyen, and Drake, Justin M.
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ANDROGEN receptors ,ABIRATERONE acetate ,CASTRATION-resistant prostate cancer ,ANDROGEN deprivation therapy ,PROTEIN-tyrosine kinases ,DNA replication ,PROSTATE cancer - Abstract
Prostate cancer (PCa) remains the most diagnosed non-skin cancer amongst the American male population. Treatment for localized prostate cancer consists of androgen deprivation therapies (ADTs), which typically inhibit androgen production and the androgen receptor (AR). Though initially effective, a subset of patients will develop resistance to ADTs and the tumors will transition to castration-resistant prostate cancer (CRPC). Second generation hormonal therapies such as abiraterone acetate and enzalutamide are typically given to men with CRPC. However, these treatments are not curative and typically prolong survival only by a few months. Several resistance mechanisms contribute to this lack of efficacy such as the emergence of AR mutations, AR amplification, lineage plasticity, AR splice variants (AR-Vs) and increased kinase signaling. Having identified SRC kinase as a key tyrosine kinase enriched in CRPC patient tumors from our previous work, we evaluated whether inhibition of SRC kinase synergizes with enzalutamide or chemotherapy in several prostate cancer cell lines expressing variable AR isoforms. We observed robust synergy between the SRC kinase inhibitor, saracatinib, and enzalutamide, in the AR-FL+/AR-V+ CRPC cell lines, LNCaP95 and 22Rv1. We also observed that saracatinib significantly decreases AR Y534 phosphorylation, a key SRC kinase substrate residue, on AR-FL and AR-Vs, along with the AR regulome, supporting key mechanisms of synergy with enzalutamide. Lastly, we also found that the saracatinib-enzalutamide combination reduced DNA replication compared to the saracatinib-docetaxel combination, resulting in marked increased apoptosis. By elucidating this combination strategy, we provide pre-clinical data that suggests combining SRC kinase inhibitors with enzalutamide in select patients that express both AR-FL and AR-Vs. [ABSTRACT FROM AUTHOR]
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- 2023
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21. ZBTB7A as a novel vulnerability in neuroendocrine prostate cancer.
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Song Yi Bae, Bergom, Hannah E., Day, Abderrahman, Greene, Joseph T., Sychev, Zoi E., Larson, Gabrianne, Corey, Eva, Plymate, Stephen R., Freedman, Tanya S., Hwang, Justin H., and Drake, Justin M.
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PROSTATE cancer ,ANDROGEN receptors ,CELL cycle ,GENE regulatory networks ,CASTRATION-resistant prostate cancer - Abstract
Neuroendocrine prostate cancer (NEPC) is a highly aggressive subtype of prostate cancer. NEPC is characterized by the loss of androgen receptor (AR) signaling and transdifferentiation toward small-cell neuroendocrine (SCN) phenotypes, which results in resistance to AR-targeted therapy. NEPC resembles other SCN carcinomas clinically, histologically and in gene expression. Here, we leveraged SCN phenotype scores of various cancer cell lines and gene depletion screens from the Cancer Dependency Map (DepMap) to identify vulnerabilities in NEPC. We discovered ZBTB7A, a transcription factor, as a candidate promoting the progression of NEPC. Cancer cells with high SCN phenotype scores showed a strong dependency on RET kinase activity with a high correlation between RET and ZBTB7A dependencies in these cells. Utilizing informatic modeling of whole transcriptome sequencing data from patient samples, we identified distinct gene networking patterns of ZBTB7A in NEPC versus prostate adenocarcinoma. Specifically, we observed a robust association of ZBTB7A with genes promoting cell cycle progression, including apoptosis regulating genes. Silencing ZBTB7A in a NEPC cell line confirmed the dependency on ZBTB7A for cell growth via suppression of the G1/S transition in the cell cycle and induction of apoptosis. Collectively, our results highlight the oncogenic function of ZBTB7A in NEPC and emphasize the value of ZBTB7A as a promising therapeutic strategy for targeting NEPC tumors. [ABSTRACT FROM AUTHOR]
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- 2023
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22. Oncogene-specific activation of tyrosine kinase networks during prostate cancer progression
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Drake, Justin M., Graham, Nicholas A., Stoyanova, Tanya, Sedghi, Amir, Goldstein, Andrew S., Cai, Houjian, Smith, Daniel A., Zhang, Hong, Komisopoulou, Evangelia, Huang, Jiaoti, Graeber, Thomas G., and Witte, Owen N.
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- 2012
23. Integrin α3β1 regulates tumor cell responses to stromal cells and can function to suppress prostate cancer metastatic colonization
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Varzavand, Afshin, Drake, Justin M., Svensson, Robert U., Herndon, Mary E., Zhou, Bo, Henry, Michael D., and Stipp, Christopher S.
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- 2013
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24. Assessing Tumor Growth and Distribution in a Model of Prostate Cancer Metastasis using Bioluminescence Imaging
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Drake, Justin M., Gabriel, Curtis L., and Henry, Michael D.
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- 2005
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25. CREB5 reprograms FOXA1 nuclear interactions to promote resistance to androgen receptor-targeting therapies.
- Author
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Hwang, Justin H., Arafeh, Rand, Ji-Heui Seo, Baca, Sylvan C., Ludwig, Megan, Arnoff, Taylor E., Sawyer, Lydia, Richter, Camden, Tape, Sydney, Bergom, Hannah E., McSweeney, Sean, Rennhack, Jonathan P., Klingenberg, Sarah A., Cheung, Alexander T. M., Kwon, Jason, So, Jonathan, Kregel, Steven, Van Allen, Eliezer M., Drake, Justin M., and Freedman, Matthew L.
- Published
- 2022
- Full Text
- View/download PDF
26. Massively parallel, computationally guided design of a proenzyme.
- Author
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Yachnin, Brahm J., Azouz, Laura R., White III, Ralph E., Minetti, Conceição A. S. A., Remeta, David P., Tan, Victor M., Drake, Justin M., and Khare, Sagar D.
- Subjects
ZYMOGENS ,INHIBITION (Chemistry) ,CATALYTIC domains ,DRUG toxicity ,COMMERCIAL products ,CELL culture - Abstract
Confining the activity of a designed protein to a specific microenvironment would have broad-ranging applications, such as enabling cell type-specific therapeutic action by enzymes while avoiding off-target effects. While many natural enzymes are synthesized as inactive zymogens that can be activated by proteolysis, it has been challenging to redesign any chosen enzyme to be similarly stimulus responsive. Here, we develop a massively parallel computational design, screening, and next-generation sequencing-based approach for proenzyme design. For a model system, we employ carboxypeptidase G2 (CPG2), a clinically approved enzyme that has applications in both the treatment of cancer and controlling drug toxicity. Detailed kinetic characterization of the most effectively designed variants shows that they are inhibited by ~80% compared to the unmodified protein, and their activity is fully restored following incubation with site-specific proteases. Introducing disulfide bonds between the pro- and catalytic domains based on the design models increases the degree of inhibition to 98% but decreases the degree of restoration of activity by proteolysis. A selected disulfide-containing proenzyme exhibits significantly lower activity relative to the fully activated enzyme when evaluated in cell culture. Structural and thermodynamic characterization provides detailed insights into the prodomain binding and inhibition mechanisms. The described methodology is general and could enable the design of a variety of proproteins with precise spatial regulation. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
27. Emerging Role of Extracellular Vesicles in Prostate Cancer.
- Author
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Ludwig, Megan, Rajvansh, Rhea, and Drake, Justin M
- Subjects
EXTRACELLULAR vesicles ,PROSTATE cancer ,PROGNOSIS ,PROSTATE ,PROSTATE-specific antigen ,METASTASIS - Abstract
Prostate cancer (PCa) is the second most common cancer among men in the United States. While the use of prostate-specific antigen has improved the ability to screen and ultimately diagnose PCa, there still remain false positives due to noncancerous conditions in the prostate gland itself and other prognostic biomarkers for PCa are needed. Contents within extracellular vesicles (EVs) have emerged as promising biomarkers that can give valuable information about disease state, and have the additional benefit of being acquired through noninvasive liquid biopsies. Meaningful communication between cancer cells and the microenvironment are carried by EVs, which impact important cellular processes in prostate cancer such as metastasis, immune regulation, and drug resistance. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
28. Nutraceutical or Pharmacological Potential of Moringa oleifera Lam.
- Author
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Kou, Xianjuan, Li, Biao, Olayanju, Julia B., Drake, Justin M., and Chen, Ning
- Abstract
Moringa oleifera Lam. (M. oleifera), which belongs to the Moringaceae family, is a perennial deciduous tropical tree, and native to the south of the Himalayan Mountains in northern India. M. oleifera is rich in proteins, vitamin A, minerals, essential amino acids, antioxidants, and flavonoids, as well as isothiocyanates. The extracts from M. oleifera exhibit multiple nutraceutical or pharmacological functions including anti-inflammatory, antioxidant, anti-cancer, hepatoprotective, neuroprotective, hypoglycemic, and blood lipid-reducing functions. The beneficial functions of M. oleifera are strongly associated with its phytochemicals such as flavonoids or isothiocyanates with bioactivity. In this review, we summarize the research progress related to the bioactivity and pharmacological mechanisms of M. oleifera in the prevention and treatment of a series of chronic diseases--including inflammatory diseases, neuro-dysfunctional diseases, diabetes, and cancers--which will provide a reference for its potential application in the prevention and treatment of chronic diseases or health promotion. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
29. Impact of Phosphoproteomics in the Era of Precision Medicine for Prostate Cancer.
- Author
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Ramroop, Johnny R., Stein, Mark N., and Drake, Justin M.
- Subjects
PROSTATE cancer ,CANCER in men ,CANCER treatment - Abstract
Prostate cancer is the most common malignancy in men in the United States. While androgen deprivation therapy results in tumor responses initially, there is relapse and progression to metastatic castration-resistant prostate cancer. Currently, all prostate cancer patients receive essentially the same treatment, and there is a need for clinically applicable technologies to provide predictive biomarkers toward personalized therapies. Genomic analyses of tumors are used for clinical applications, but with a paucity of obvious driver mutations in metastatic castration-resistant prostate cancer, other applications, such as phosphoproteomics, may complement this approach. Immunohistochemistry and reverse phase protein arrays are limited by the availability of reliable antibodies and evaluates a preselected number of targets. Mass spectrometry-based phosphoproteomics has been used to profile tumors consisting of thousands of phosphopeptides from individual patients after surgical resection or at autopsy. However, this approach is time consuming, and while a large number of candidate phosphopeptides are obtained for evaluation, limitations are reduced reproducibility, sensitivity, and precision. Targeted mass spectrometry can help eliminate these limitations and is more cost effective and less time consuming making it a practical platform for future clinical testing. In this review, we discuss the use of phosphoproteomics in prostate cancer and other clinical cancer tissues for target identification, hypothesis testing, and possible patient stratification. We highlight the majority of studies that have used phosphoproteomics in prostate cancer tissues and cell lines and propose ways forward to apply this approach in basic and clinical research. Overall, the implementation of phosphoproteomics via targeted mass spectrometry has tremendous potential to aid in the development of more rational, personalized therapies that will result in increased survival and quality of life enhancement in patients suffering from metastatic castration-resistant prostate cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
30. Bone-specific growth inhibition of prostate cancer metastasis by atrasentan.
- Author
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Drake, Justin M., Danke, Joshua R., and Henry, Michael D.
- Published
- 2010
- Full Text
- View/download PDF
31. Regulated proteolysis of Trop2 drives epithelial hyperplasia and stem cell self-renewal via β-catenin signaling.
- Author
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Stoyanova, Tanya, Goldstein, Andrew S., Cai, Houjian, Drake, Justin M., Jiaoti Huang, and Witte, Owen N.
- Subjects
- *
CELL membranes , *PROTEOLYSIS , *PROTEIN metabolism , *NEOPLASTIC cell transformation , *CATENINS , *STEM cell culture - Abstract
The cell surface protein Trop2 is expressed on immature stem/progenitor-like cells and is overexpressed in many epithelial cancers. However the biological function of Trop2 in tissue maintenance and tumorigenesis remains unclear. In this study, we demonstrate that Trop2 is a regulator of self-renewal, proliferation, and transformation. Trop2 controls these processes through a mechanism of regulated intramembrane proteolysis that leads to cleavage of Trop2, creating two products: the extracellular domain and the intracellular domain. The intracellular domain of Trop2 is released from the membrane and accumulates in the nucleus. Heightened expression of the Trop2 intracellular domain promotes stem/progenitor self-renewal through signaling via β-catenin and is sufficient to initiate precursor lesions to prostate cancer in vivo. Importantly, we demonstrate that loss of β-catenin or Trop2 loss-of-function cleavage mutants abrogates Trop2-driven self-renewal and hyperplasia in the prostate. These findings suggest that heightened expression of Trop2 is selected for in epithelial cancers to enhance the stem-like properties of self-renewal and proliferation. Defining the mechanism of Trop2 function in self-renewal and transformation is essential to identify new therapeutic strategies to block Trop2 activation in cancer. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
32. Dual inhibition of ATR and DNA-PKcs radiosensitizes ATM-mutant prostate cancer.
- Author
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Hofstad M, Woods A, Parra K, Sychev ZE, Mazzagatti A, Yu L, Gilbreath C, Ly P, Drake JM, and Kittler R
- Abstract
In advanced castration resistant prostate cancer (CRPC), mutations in the DNA damage response (DDR) gene ataxia telangiectasia mutated ( ATM ) are common. While poly(ADP-ribose) polymerase inhibitors are approved in this context, their clinical efficacy remains limited. Thus, there is a compelling need to identify alternative therapeutic avenues for ATM mutant prostate cancer patients. Here, we generated matched ATM-proficient and ATM-deficient CRPC lines to elucidate the impact of ATM loss on DDR in response to DNA damage via irradiation. Through unbiased phosphoproteomic screening, we unveiled that ATM-deficient CRPC lines maintain dependence on downstream ATM targets through activation of ATR and DNA-PKcs kinases. Dual inhibition of ATR and DNA-PKcs effectively inhibited downstream γH2AX foci formation in response to irradiation and radiosensitized ATM-deficient lines to a greater extent than either ATM-proficient controls or single drug treatment. Further, dual inhibition abrogated residual downstream ATM pathway signaling and impaired replication fork dynamics. To circumvent potential toxicity, we leveraged the RUVBL1/2 ATPase inhibitor Compound B, which leads to the degradation of both ATR and DNA-PKcs kinases. Compound B effectively radiosensitized ATM-deficient CRPC in vitro and in vivo , and impacted replication fork dynamics. Overall, dual targeting of both ATR and DNA-PKcs is necessary to block DDR in ATM-deficient CRPC, and Compound B could be utilized as a novel therapy in combination with irradiation in these patients.
- Published
- 2024
- Full Text
- View/download PDF
33. Unraveling the Global Proteome and Phosphoproteome of Prostate Cancer Patient-Derived Xenografts.
- Author
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Sychev ZE, Day A, Bergom HE, Larson G, Ali A, Ludwig M, Boytim E, Coleman I, Corey E, Plymate SR, Nelson PS, Hwang JH, and Drake JM
- Subjects
- Humans, Male, Animals, Mice, Prostatic Neoplasms, Castration-Resistant metabolism, Prostatic Neoplasms, Castration-Resistant genetics, Prostatic Neoplasms, Castration-Resistant pathology, Heterografts, Prostatic Neoplasms metabolism, Prostatic Neoplasms genetics, Prostatic Neoplasms pathology, Adenocarcinoma metabolism, Adenocarcinoma genetics, Adenocarcinoma pathology, Xenograft Model Antitumor Assays, Proteomics methods, Proteome metabolism, Phosphoproteins metabolism
- Abstract
Resistance to androgen-deprivation therapies leads to metastatic castration-resistant prostate cancer (mCRPC) of adenocarcinoma (AdCa) origin that can transform into emergent aggressive variant prostate cancer (AVPC), which has neuroendocrine (NE)-like features. In this work, we used LuCaP patient-derived xenograft (PDX) tumors, clinically relevant models that reflect and retain key features of the tumor from advanced prostate cancer patients. Here we performed proteome and phosphoproteome characterization of 48 LuCaP PDX tumors and identified over 94,000 peptides and 9,700 phosphopeptides corresponding to 7,738 proteins. We compared 15 NE versus 33 AdCa samples, which included six different PDX tumors for each group in biological replicates, and identified 309 unique proteins and 476 unique phosphopeptides that were significantly altered and corresponded to proteins that are known to distinguish these two phenotypes. Assessment of concordance from PDX tumor-matched protein and mRNA revealed increased dissonance in transcriptionally regulated proteins in NE and metabolite interconversion enzymes in AdCa., Implications: Overall, our study highlights the importance of protein-based identification when compared with RNA and provides a rich resource of new and feasible targets for clinical assay development and in understanding the underlying biology of these tumors., (©2024 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2024
- Full Text
- View/download PDF
34. Adipose Triglyceride Lipase Is a Therapeutic Target in Advanced Prostate Cancer That Promotes Metabolic Plasticity.
- Author
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Awad D, Cao PHA, Pulliam TL, Spradlin M, Subramani E, Tellman TV, Ribeiro CF, Muzzioli R, Jewell BE, Pakula H, Ackroyd JJ, Murray MM, Han JJ, Leng M, Jain A, Piyarathna B, Liu J, Song X, Zhang J, Klekers AR, Drake JM, Ittmann MM, Coarfa C, Piwnica-Worms D, Farach-Carson MC, Loda M, Eberlin LS, and Frigo DE
- Subjects
- Male, Humans, Mice, Animals, Lipolysis genetics, Lipid Metabolism, Lipase genetics, Lipase metabolism, Serine metabolism, Tumor Microenvironment, Calcium-Calmodulin-Dependent Protein Kinase Kinase, Prostatic Neoplasms, Castration-Resistant
- Abstract
Lipid metabolism plays a central role in prostate cancer. To date, the major focus has centered on de novo lipogenesis and lipid uptake in prostate cancer, but inhibitors of these processes have not benefited patients. A better understanding of how cancer cells access lipids once they are created or taken up and stored could uncover more effective strategies to perturb lipid metabolism and treat patients. Here, we identified that expression of adipose triglyceride lipase (ATGL), an enzyme that controls lipid droplet homeostasis and a previously suspected tumor suppressor, correlates with worse overall survival in men with advanced, castration-resistant prostate cancer (CRPC). Molecular, genetic, or pharmacologic inhibition of ATGL impaired human and murine prostate cancer growth in vivo and in cell culture or organoids under conditions mimicking the tumor microenvironment. Mass spectrometry imaging demonstrated that ATGL profoundly regulates lipid metabolism in vivo, remodeling membrane composition. ATGL inhibition induced metabolic plasticity, causing a glycolytic shift that could be exploited therapeutically by cotargeting both metabolic pathways. Patient-derived phosphoproteomics identified ATGL serine 404 as a target of CAMKK2-AMPK signaling in CRPC cells. Mutation of serine 404 did not alter the lipolytic activity of ATGL but did decrease CRPC growth, migration, and invasion, indicating that noncanonical ATGL activity also contributes to disease progression. Unbiased immunoprecipitation/mass spectrometry suggested that mutation of serine 404 not only disrupts existing ATGL protein interactions but also leads to new protein-protein interactions. Together, these data nominate ATGL as a therapeutic target for CRPC and provide insights for future drug development and combination therapies., Significance: ATGL promotes prostate cancer metabolic plasticity and progression through both lipase-dependent and lipase-independent activity, informing strategies to target ATGL and lipid metabolism for cancer treatment., (©2023 American Association for Cancer Research.)
- Published
- 2024
- Full Text
- View/download PDF
35. Unraveling the Global Proteome and Phosphoproteome of Prostate Cancer Patient-Derived Xenografts.
- Author
-
Sychev ZE, Day A, Bergom HE, Larson G, Ali A, Ludwig M, Boytim E, Coleman I, Corey E, Plymate SR, Nelson PS, Hwang JH, and Drake JM
- Abstract
Resistance to androgen deprivation therapies leads to metastatic castration-resistant prostate cancer (mCRPC) of adenocarcinoma (AdCa) origin that can transform to emergent aggressive variant prostate cancer (AVPC) which has neuroendocrine (NE)-like features. To this end, we used LuCaP patient-derived xenograft (PDX) tumors, clinically relevant models that reflects and retains key features of the tumor from advanced prostate cancer patients. Here we performed proteome and phosphoproteome characterization of 48 LuCaP PDX tumors and identified over 94,000 peptides and 9,700 phosphopeptides corresponding to 7,738 proteins. When we compared 15 NE versus 33 AdCa PDX samples, we identified 309 unique proteins and 476 unique phosphopeptides that were significantly altered and corresponded to proteins that are known to distinguish these two phenotypes. Assessment of protein and RNA concordance from these tumors revealed increased dissonance in transcriptionally regulated proteins in NE and metabolite interconversion enzymes in AdCa., Competing Interests: Conflict of Interest Statement ZES has no conflicts relevant to this work, however she works as a consultant to Astrin Bioscience as a senior scientist. EC is a consultant of DotQuant, and obtained research support form AbbVie, Genentech, Bayer Pharmaceuticals, Forma Therapeutics, KronoBio, Foghorn, MacroGenics, Gilead, Janssen Research and GSK. . JMD has no conflicts relevant to this work. However, he holds equity in and serves as Chief Scientific Officer of Astrin Biosciences. This interest has been reviewed and managed by the University of Minnesota in accordance with its Conflict-of-Interest policies. None of these companies contributed to or directed any of the research reported in this article. The remaining authors declare no potential conflicts of interest.
- Published
- 2023
- Full Text
- View/download PDF
36. Saracatinib synergizes with enzalutamide to downregulate AR activity in CRPC.
- Author
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White RE 3rd, Bannister M, Day A, Bergom HE, Tan VM, Hwang J, Dang Nguyen H, and Drake JM
- Abstract
Prostate cancer (PCa) remains the most diagnosed non-skin cancer amongst the American male population. Treatment for localized prostate cancer consists of androgen deprivation therapies (ADTs), which typically inhibit androgen production and the androgen receptor (AR). Though initially effective, a subset of patients will develop resistance to ADTs and the tumors will transition to castration-resistant prostate cancer (CRPC). Second generation hormonal therapies such as abiraterone acetate and enzalutamide are typically given to men with CRPC. However, these treatments are not curative and typically prolong survival only by a few months. Several resistance mechanisms contribute to this lack of efficacy such as the emergence of AR mutations, AR amplification, lineage plasticity, AR splice variants (AR-Vs) and increased kinase signaling. Having identified SRC kinase as a key tyrosine kinase enriched in CRPC patient tumors from our previous work, we evaluated whether inhibition of SRC kinase synergizes with enzalutamide or chemotherapy in several prostate cancer cell lines expressing variable AR isoforms. We observed robust synergy between the SRC kinase inhibitor, saracatinib, and enzalutamide, in the AR-FL+/AR-V+ CRPC cell lines, LNCaP95 and 22Rv1. We also observed that saracatinib significantly decreases AR Y
534 phosphorylation, a key SRC kinase substrate residue, on AR-FL and AR-Vs, along with the AR regulome, supporting key mechanisms of synergy with enzalutamide. Lastly, we also found that the saracatinib-enzalutamide combination reduced DNA replication compared to the saracatinib-docetaxel combination, resulting in marked increased apoptosis. By elucidating this combination strategy, we provide pre-clinical data that suggests combining SRC kinase inhibitors with enzalutamide in select patients that express both AR-FL and AR-Vs., Competing Interests: JD has no conflicts relevant to this work. However, he holds equity in and serves as Chief Scientific Officer of Astrin Biosciences. This interest has been reviewed and managed by the University of Minnesota in accordance with its Conflict of Interest policies. None of these companies contributed to or directed any of the research reported in this article. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest., (Copyright © 2023 White, Bannister, Day, Bergom, Tan, Hwang, Dang Nguyen and Drake.)- Published
- 2023
- Full Text
- View/download PDF
37. Saracatinib synergizes with enzalutamide to downregulate androgen receptor activity in castration resistant prostate cancer.
- Author
-
White RE, Bannister M, Day A, Bergom HE, Tan VM, Hwang J, Nguyen HD, and Drake JM
- Abstract
Prostate cancer (PCa) remains the most diagnosed non-skin cancer amongst the American male population. Treatment for localized prostate cancer consists of androgen deprivation therapies (ADTs), which typically inhibit androgen production and the androgen receptor (AR). Though initially effective, a subset of patients will develop resistance to ADTs and the tumors will transition to castration-resistant prostate cancer (CRPC). Second generation hormonal therapies such as abiraterone acetate and enzalutamide are typically given to men with CRPC. However, these treatments are not curative and typically prolong survival only by a few months. Several resistance mechanisms contribute to this lack of efficacy such as the emergence of AR mutations, AR amplification, lineage plasticity, AR splice variants (AR-Vs) and increased kinase signaling. Having identified SRC kinase as a key tyrosine kinase enriched in CRPC patient tumors from our previous work, we evaluated whether inhibition of SRC kinase synergizes with enzalutamide or chemotherapy in several prostate cancer cell lines expressing variable AR isoforms. We observed robust synergy between the SRC kinase inhibitor, saracatinib, and enzalutamide, in the AR-FL+/AR-V+ CRPC cell lines, LNCaP95 and 22Rv1. We also observed that saracatinib significantly decreases AR Y
534 phosphorylation, a key SRC kinase substrate residue, on AR-FL and AR-Vs, along with the AR regulome, supporting key mechanisms of synergy with enzalutamide. Lastly, we also found that the saracatinib-enzalutamide combination reduced DNA replication compared to the saracatinib-docetaxel combination, resulting in marked increased apoptosis. By elucidating this combination strategy, we provide pre-clinical data that suggests combining SRC kinase inhibitors with enzalutamide in select patients that express both AR-FL and AR-Vs.- Published
- 2023
- Full Text
- View/download PDF
38. Induction of PARP7 Creates a Vulnerability for Growth Inhibition by RBN2397 in Prostate Cancer Cells.
- Author
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Yang C, Wierbiłowicz K, Dworak NM, Bae SY, Tengse SB, Abianeh N, Drake JM, Abbas T, Ratan A, Wotton D, and Paschal BM
- Subjects
- Male, Humans, Prostate metabolism, ADP Ribose Transferases genetics, Androgens, Receptors, Androgen genetics, Prostatic Neoplasms drug therapy
- Abstract
The ADP-ribosyltransferase PARP7 modulates protein function by conjugating ADP-ribose to the side chains of acceptor amino acids. PARP7 has been shown to affect gene expression in prostate cancer cells and certain other cell types by mechanisms that include transcription factor ADP-ribosylation. Here, we use a recently developed catalytic inhibitor to PARP7, RBN2397, to study the effects of PARP7 inhibition in androgen receptor (AR)-positive and AR-negative prostate cancer cells. We find that RBN2397 has nanomolar potency for inhibiting androgen-induced ADP-ribosylation of the AR. RBN2397 inhibits the growth of prostate cancer cells in culture when cells are treated with ligands that activate the AR, or the aryl hydrocarbon receptor, and induce PARP7 expression. We show that the growth-inhibitory effects of RBN2397 are distinct from its enhancement of IFN signaling recently shown to promote tumor immunogenicity. RBN2397 treatment also induces trapping of PARP7 in a detergent-resistant fraction within the nucleus, which is reminiscent of how inhibitors such as talazoparib affect PARP1 compartmentalization. Because PARP7 is expressed in AR-negative metastatic tumors and RBN2397 can affect cancer cells through multiple mechanisms, PARP7 may be an actionable target in advanced prostate cancer., Significance: RBN2397 is a potent and selective inhibitor of PARP7 that reduces the growth of prostate cancer cells, including a model for treatment-emergent neuroendocrine prostate cancer. RBN2397 induces PARP7 trapping on chromatin, suggesting its mechanism of action might be similar to clinically used PARP1 inhibitors., (© 2023 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2023
- Full Text
- View/download PDF
39. ZBTB7A as a novel vulnerability in neuroendocrine prostate cancer.
- Author
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Bae SY, Bergom HE, Day A, Greene JT, Sychev ZE, Larson G, Corey E, Plymate SR, Freedman TS, Hwang JH, and Drake JM
- Subjects
- Humans, Male, Transcription Factors metabolism, DNA-Binding Proteins genetics, Cell Line, Tumor, Prostatic Neoplasms pathology, Neuroendocrine Tumors genetics, Neuroendocrine Tumors pathology
- Abstract
Neuroendocrine prostate cancer (NEPC) is a highly aggressive subtype of prostate cancer. NEPC is characterized by the loss of androgen receptor (AR) signaling and transdifferentiation toward small-cell neuroendocrine (SCN) phenotypes, which results in resistance to AR-targeted therapy. NEPC resembles other SCN carcinomas clinically, histologically and in gene expression. Here, we leveraged SCN phenotype scores of various cancer cell lines and gene depletion screens from the Cancer Dependency Map (DepMap) to identify vulnerabilities in NEPC. We discovered ZBTB7A, a transcription factor, as a candidate promoting the progression of NEPC. Cancer cells with high SCN phenotype scores showed a strong dependency on RET kinase activity with a high correlation between RET and ZBTB7A dependencies in these cells. Utilizing informatic modeling of whole transcriptome sequencing data from patient samples, we identified distinct gene networking patterns of ZBTB7A in NEPC versus prostate adenocarcinoma. Specifically, we observed a robust association of ZBTB7A with genes promoting cell cycle progression, including apoptosis regulating genes. Silencing ZBTB7A in a NEPC cell line confirmed the dependency on ZBTB7A for cell growth via suppression of the G1/S transition in the cell cycle and induction of apoptosis. Collectively, our results highlight the oncogenic function of ZBTB7A in NEPC and emphasize the value of ZBTB7A as a promising therapeutic strategy for targeting NEPC tumors., Competing Interests: EC is consultant of Dotquant and received research funding via institutional SRAs from AbbVie, Janssen Research, Gilead, Zenith Epigenetics, Bayer, GSK, Astra Zeneca, Kronos, Foghorn, MacroGenics, and Forma Pharmaceuticals. JMD has no conflicts relevant to this work. However, he holds equity in and serves as Chief Scientific Officer of Astrin Biosciences. This interest has been reviewed and managed by the University of Minnesota in accordance with its Conflict of Interest policies. None of these companies contributed to or directed any of the research reported in this article. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Bae, Bergom, Day, Greene, Sychev, Larson, Corey, Plymate, Freedman, Hwang and Drake.)
- Published
- 2023
- Full Text
- View/download PDF
40. Unique-region phosphorylation targets LynA for rapid degradation, tuning its expression and signaling in myeloid cells.
- Author
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Brian BF 4th, Jolicoeur AS, Guerrero CR, Nunez MG, Sychev ZE, Hegre SA, Sætrom P, Habib N, Drake JM, Schwertfeger KL, and Freedman TS
- Subjects
- Animals, Humans, Jurkat Cells, Mice, Knockout, Phosphorylation, Proteolysis, Proto-Oncogene Proteins c-cbl genetics, Ubiquitin metabolism, src-Family Kinases genetics, Macrophages metabolism, Mast Cells metabolism, Myeloid Cells metabolism, Proto-Oncogene Proteins c-cbl metabolism, src-Family Kinases metabolism
- Abstract
The activity of Src-family kinases (SFKs), which phosphorylate immunoreceptor tyrosine-based activation motifs (ITAMs), is a critical factor regulating myeloid-cell activation. We reported previously that the SFK LynA is uniquely susceptible to rapid ubiquitin-mediated degradation in macrophages, functioning as a rheostat regulating signaling (Freedman et al., 2015). We now report the mechanism by which LynA is preferentially targeted for degradation and how cell specificity is built into the LynA rheostat. Using genetic, biochemical, and quantitative phosphopeptide analyses, we found that the E3 ubiquitin ligase c-Cbl preferentially targets LynA via a phosphorylated tyrosine (Y32) in its unique region. This distinct mode of c-Cbl recognition depresses steady-state expression of LynA in macrophages derived from mice. Mast cells, however, express little c-Cbl and have correspondingly high LynA. Upon activation, mast-cell LynA is not rapidly degraded, and SFK-mediated signaling is amplified relative to macrophages. Cell-specific c-Cbl expression thus builds cell specificity into the LynA checkpoint., Competing Interests: BB, AJ, CG, MN, ZS, SH, PS, NH, JD, KS, TF No competing interests declared, (© 2019, Brian et al.)
- Published
- 2019
- Full Text
- View/download PDF
41. MAPK Reliance via Acquired CDK4/6 Inhibitor Resistance in Cancer.
- Author
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de Leeuw R, McNair C, Schiewer MJ, Neupane NP, Brand LJ, Augello MA, Li Z, Cheng LC, Yoshida A, Courtney SM, Hazard ES, Hardiman G, Hussain MH, Diehl JA, Drake JM, Kelly WK, and Knudsen KE
- Subjects
- Cell Cycle Checkpoints drug effects, Cell Line, Tumor, Cell Movement drug effects, Cell Proliferation drug effects, Cyclin-Dependent Kinase 4 antagonists & inhibitors, Cyclin-Dependent Kinase 6 antagonists & inhibitors, Drug Resistance, Neoplasm drug effects, Dual Specificity Phosphatase 1 antagonists & inhibitors, Humans, MAP Kinase Kinase 1 antagonists & inhibitors, MAP Kinase Kinase 1 genetics, MAP Kinase Kinase Kinases antagonists & inhibitors, Neoplasm Invasiveness genetics, Neoplasm Invasiveness pathology, Neoplasms genetics, Neoplasms pathology, Phosphorylation drug effects, Piperazines pharmacology, Pyridines pharmacology, Retinoblastoma Protein genetics, Sequence Analysis, RNA, Signal Transduction drug effects, Xenograft Model Antitumor Assays, Cyclin-Dependent Kinase 4 genetics, Cyclin-Dependent Kinase 6 genetics, Dual Specificity Phosphatase 1 genetics, MAP Kinase Kinase Kinases genetics, Neoplasms drug therapy
- Abstract
Purpose: Loss of cell-cycle control is a hallmark of cancer, which can be targeted with agents, including cyclin-dependent kinase-4/6 (CDK4/6) kinase inhibitors that impinge upon the G
1 -S cell-cycle checkpoint via maintaining activity of the retinoblastoma tumor suppressor (RB). This class of drugs is under clinical investigation for various solid tumor types and has recently been FDA-approved for treatment of breast cancer. However, development of therapeutic resistance is not uncommon. Experimental Design: In this study, palbociclib (a CDK4/6 inhibitor) resistance was established in models of early stage, RB-positive cancer. Results: This study demonstrates that acquired palbociclib resistance renders cancer cells broadly resistant to CDK4/6 inhibitors. Acquired resistance was associated with aggressive in vitro and in vivo phenotypes, including proliferation, migration, and invasion. Integration of RNA sequencing analysis and phosphoproteomics profiling revealed rewiring of the kinome, with a strong enrichment for enhanced MAPK signaling across all resistance models, which resulted in aggressive in vitro and in vivo phenotypes and prometastatic signaling. However, CDK4/6 inhibitor-resistant models were sensitized to MEK inhibitors, revealing reliance on active MAPK signaling to promote tumor cell growth and invasion. Conclusions: In sum, these studies identify MAPK reliance in acquired CDK4/6 inhibitor resistance that promotes aggressive disease, while nominating MEK inhibition as putative novel therapeutic strategy to treat or prevent CDK4/6 inhibitor resistance in cancer. Clin Cancer Res; 24(17); 4201-14. ©2018 AACR ., (©2018 American Association for Cancer Research.)- Published
- 2018
- Full Text
- View/download PDF
42. Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer.
- Author
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Cheng LC, Li Z, Graeber TG, Graham NA, and Drake JM
- Subjects
- Humans, Male, Prostatic Neoplasms metabolism, Prostatic Neoplasms pathology, Phosphopeptides analysis, Prostatic Neoplasms genetics, Proteomics methods, Tandem Mass Spectrometry methods
- Abstract
Phosphoproteomics involves the large-scale study of phosphorylated proteins. Protein phosphorylation is a critical step in many signal transduction pathways and is tightly regulated by kinases and phosphatases. Therefore, characterizing the phosphoproteome may provide insights into identifying novel targets and biomarkers for oncologic therapy. Mass spectrometry provides a way to globally detect and quantify thousands of unique phosphorylation events. However, phosphopeptides are much less abundant than non-phosphopeptides, making biochemical analysis more challenging. To overcome this limitation, methods to enrich phosphopeptides prior to the mass spectrometry analysis are required. We describe a procedure to extract and digest proteins from tissue to yield peptides, followed by an enrichment for phosphotyrosine (pY) and phosphoserine/threonine (pST) peptides using an antibody-based and/or titanium dioxide (TiO2)-based enrichment method. After the sample preparation and mass spectrometry, we subsequently identify and quantify phosphopeptides using liquid chromatography-mass spectrometry and analysis software.
- Published
- 2018
- Full Text
- View/download PDF
43. Gain-of-function mutant p53 activates small GTPase Rac1 through SUMOylation to promote tumor progression.
- Author
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Yue X, Zhang C, Zhao Y, Liu J, Lin AW, Tan VM, Drake JM, Liu L, Boateng MN, Li J, Feng Z, and Hu W
- Subjects
- Cell Line, Cell Line, Tumor, Cysteine Endopeptidases metabolism, Disease Progression, Humans, Neoplasm Metastasis, Neoplasms enzymology, Neoplasms metabolism, Neoplasms pathology, Tumor Suppressor Protein p53 metabolism, Carcinogenesis genetics, Mutation, Sumoylation, Tumor Suppressor Protein p53 genetics, rac1 GTP-Binding Protein metabolism
- Abstract
Tumor suppressor p53 is frequently mutated in human cancer. Mutant p53 often promotes tumor progression through gain-of-function (GOF) mechanisms. However, the mechanisms underlying mutant p53 GOF are not well understood. In this study, we found that mutant p53 activates small GTPase Rac1 as a critical mechanism for mutant p53 GOF to promote tumor progression. Mechanistically, mutant p53 interacts with Rac1 and inhibits its interaction with SUMO-specific protease 1 (SENP1), which in turn inhibits SENP1-mediated de-SUMOylation of Rac1 to activate Rac1. Targeting Rac1 signaling by RNAi, expression of the dominant-negative Rac1 (Rac1 DN), or the specific Rac1 inhibitor NSC23766 greatly inhibits mutant p53 GOF in promoting tumor growth and metastasis. Furthermore, mutant p53 expression is associated with enhanced Rac1 activity in clinical tumor samples. These results uncover a new mechanism for Rac1 activation in tumors and, most importantly, reveal that activation of Rac1 is an unidentified and critical mechanism for mutant p53 GOF in tumorigenesis, which could be targeted for therapy in tumors containing mutant p53., (© 2017 Yue et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2017
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44. Complementing genomics and transcriptomics: Phosphoproteomics illuminates systems biology in prostate cancer.
- Author
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Tan VM, Cheng LC, and Drake JM
- Abstract
Integration of phosphoproteomics with traditional genomics and transcriptomics provides a more comprehensive overview of the signaling networks in advanced prostate cancer for immediate preclinical and future clinical use. Our recent publication introduces computational approaches for integrating the phosphoproteome, specifically with the intent of identifying important kinase signaling networks in advanced-stage prostate cancer.
- Published
- 2016
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45. Purification and direct transformation of epithelial progenitor cells from primary human prostate.
- Author
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Goldstein AS, Drake JM, Burnes DL, Finley DS, Zhang H, Reiter RE, Huang J, and Witte ON
- Subjects
- Animals, Cells, Cultured, Humans, Male, Mice, Cell Transformation, Neoplastic, Epithelial Cells pathology, Prostate cytology, Prostatic Neoplasms pathology, Stem Cells pathology
- Abstract
Epithelial cell transformation has been demonstrated in numerous animal models for the study of solid tumor biology. However, little evidence exists for human epithelial cell transformation without previous immortalization via genetic influences such as SV40 T-antigen, thus limiting our knowledge of the events that can transform naive human epithelium. Here we describe a system developed in our laboratory to directly transform freshly isolated primary human prostate epithelial cells without previous culture or immortalization. Prostate tissue is obtained from patients and benign tissue is separated from malignant tissue. Benign and malignant tissues are mechanically and enzymatically dissociated to single cells overnight, and immune cells and epithelial subsets are isolated on the basis of differential expression of surface antigens. Epithelial progenitor cells are transduced with lentiviruses expressing oncogenes and combined with inductive stroma for in vivo studies. At 8-16 weeks after transplantation into immune-deficient mice, the development of lesions, histologically classified as benign prostate, prostatic intraepithelial neoplasia and adenocarcinoma, can be evaluated.
- Published
- 2011
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- View/download PDF
46. ZEB1 coordinately regulates laminin-332 and {beta}4 integrin expression altering the invasive phenotype of prostate cancer cells.
- Author
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Drake JM, Barnes JM, Madsen JM, Domann FE, Stipp CS, and Henry MD
- Subjects
- Cell Line, Tumor, Cell Movement, Chromatin metabolism, Coculture Techniques, Epithelial Cells cytology, Extracellular Matrix metabolism, Humans, Male, Neoplasm Metastasis, Phenotype, Zinc Finger E-box-Binding Homeobox 1, Gene Expression Regulation, Neoplastic, Homeodomain Proteins metabolism, Integrin beta4 metabolism, Laminin chemistry, Prostatic Neoplasms metabolism, Transcription Factors metabolism
- Abstract
Metastasis involves the invasion of cancer cells across both the extracellular matrix and cellular barriers, and an evolving theme is that epithelial-to-mesenchymal transition (EMT) may mediate invasive cellular behavior. Previously, we isolated and analyzed a subpopulation of PC-3 prostate cancer cells, TEM4-18, and found that these cells both invaded an endothelial barrier more efficiently and exhibited enhanced metastatic colonization in vivo. Transendothelial migration of these cells depended on expression of ZEB1, a known regulator of EMT. Surprisingly, these cells were much less invasive than parental PC-3 cells in assays that involve matrix barriers. Here, we report that TEM4-18 cells express significantly reduced levels of two subunits of laminin-332 (β3 and γ2) and that exogenous laminin-332, or co-culture with laminin-332-expressing cells, rescues the in vitro invasion phenotype in these cells. Stable knockdown of ZEB1 in prostate cancer cells up-regulated LAMC2 and ITGB4 mRNA and protein and resulted in a concomitant increase in Transwell migration. Using chromatin immunoprecipitation (ChIP), we show that ZEB1 directly interacts with the promoters of LAMC2 and ITGB4. These results provide a novel molecular basis for reduced laminin-332 observed in clinical prostate cancer specimens and demonstrate a context-dependent role for EMT in invasive cellular behavior.
- Published
- 2010
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- View/download PDF
47. ZEB1 enhances transendothelial migration and represses the epithelial phenotype of prostate cancer cells.
- Author
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Drake JM, Strohbehn G, Bair TB, Moreland JG, and Henry MD
- Subjects
- Animals, Cadherins metabolism, Cell Line, Tumor, Endothelial Cells metabolism, Epithelial Cells metabolism, Epithelium pathology, Gene Expression Regulation, Neoplastic, Homeodomain Proteins genetics, Humans, Male, Mesoderm pathology, Mice, Neoplasm Metastasis, Oligonucleotide Array Sequence Analysis, Phenotype, Prostatic Neoplasms genetics, Transcription Factors genetics, Xenograft Model Antitumor Assays, Zinc Finger E-box-Binding Homeobox 1, Cell Movement, Endothelial Cells pathology, Epithelial Cells pathology, Homeodomain Proteins metabolism, Prostatic Neoplasms metabolism, Prostatic Neoplasms pathology, Transcription Factors metabolism
- Abstract
Metastatic colonization involves cancer cell lodgment or adherence in the microvasculature and subsequent migration of those cells across the endothelium into a secondary organ site. To study this process further, we analyzed transendothelial migration of human PC-3 prostate cancer cells in vitro. We isolated a subpopulation of cells, TEM4-18, that crossed an endothelial barrier more efficiently, but surprisingly, were less invasive than parental PC-3 cells in other contexts in vitro. Importantly, TEM4-18 cells were more aggressive than PC-3 cells in a murine metastatic colonization model. Microarray and FACS analysis of these cells showed that the expression of many genes previously associated with leukocyte trafficking and cancer cell extravasation were either unchanged or down-regulated. Instead, TEM4-18 cells exhibited characteristic molecular markers of an epithelial-to-mesenchymal transition (EMT), including frank loss of E-cadherin expression and up-regulation of the E-cadherin repressor ZEB1. Silencing ZEB1 in TEM4-18 cells resulted in increased E-cadherin and reduced transendothelial migration. TEM4-18 cells also express N-cadherin, which was found to be necessary, but not sufficient for increased transendothelial migration. Our results extend the role of EMT in metastasis to transendothelial migration and implicate ZEB1 and N-cadherin in this process in prostate cancer cells.
- Published
- 2009
- Full Text
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48. Endothelin-1 inhibits prostate cancer growth in vivo through vasoconstriction of tumor-feeding arterioles.
- Author
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Weydert CJ, Esser AK, Mejia RA, Drake JM, Barnes JM, and Henry MD
- Subjects
- Animals, Arterioles metabolism, Male, Mice, Arterioles pathology, Endothelin-1 physiology, Neovascularization, Pathologic prevention & control, Prostatic Neoplasms blood supply, Vasoconstriction
- Abstract
The vasoactive peptide endothelin-1 (ET-1) has been implicated in promoting the progression of prostate and other cancers though its precise mechanism(s)-of-action remain unclear. To better define the role of ET-1 in prostate cancer progression, we generated prostate cancer cell lines (PC-3 and 22Rv1) that express elevated levels of ET-1. As anticipated, increased ET-1 lead to modest autocrine growth stimulation of PC-3 cells in monolayer culture and increased colony formation in soft agar by both cell lines. Unexpectedly, however, metastatic colonization of 22Rv1 cells expressing elevated levels of ET-1 was reduced, as was the size of subcutaneous tumors produced by both 22Rv1- and PC-3 cells. Based on these data, we hypothesized that high levels of ET-1 may negatively impact the tumor microenvironment. We found that increased ET-1 expression did not consistently inhibit angiogenesis, indicating that this was not the cause of poor tumor growth. As an alternative explanation, we examined whether elevated ET-1 results in local vasoconstriction and thus reduces the blood supply available to the tumor. Consistent with this hypothesis, treatment of mice bearing PC-3 xenografts with a vasodilator increased tumor perfusion and partially restored tumor growth. Moreover, analysis of tumor vascular casts indicated vasoconstriction of tumor-feeding arterioles. Taken together, our data suggest that the local concentration of the ET-1 peptide is critical for determining a balance between its previously unrecognized tumor growth-suppressing activity (vasoconstriction) and known growth-promoting (mitogenesis, survival and angiogenesis) activities. These findings may have implications for the modification of current prostate cancer therapies involving ET-1.
- Published
- 2009
- Full Text
- View/download PDF
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