36 results on '"Jean J. Zhao"'
Search Results
2. Targeting tumor-associated macrophages with STING agonism improves the antitumor efficacy of osimertinib in a mouse model of EGFR-mutant lung cancer
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Ziying Lin, Qiwei Wang, Tao Jiang, Weihua Wang, and Jean J. Zhao
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lung cancer ,EGFR-mutant ,Osimertinib ,STING agonist ,tumor-associated macrophages ,Immunologic diseases. Allergy ,RC581-607 - Abstract
IntroductionDespite the impressive clinical response rate of osimertinib, a third-generation EGFR-TKI, as a frontline treatment for patients with EGFR-mutant non-small-cell lung cancer (NSCLC) or as a salvage therapy for patients with T790M mutation, resistance to osimertinib is common in the clinic. The mechanisms underlying osimertinib resistance are heterogenous. While genetic mutations within EGFR or other cancer driver pathways mediated mechanisms are well-documented, the role of tumor cell and tumor immune microenvironment in mediating the response to osimertinib remains elusive.Methods and resultsHere, using a syngeneic mouse model of EGFR-mutant lung cancer, we show that tumor regression elicited by osimertinib requires activation of CD8+ T cells. However, tumor-associated macrophages (TAMs) accumulated in advanced tumors inhibit CD8+ T cell activation and diminish the response to osimertinib. These results are corroborated by analyses of clinical data. Notably, reprogramming TAMs with a systemic STING agonist MSA-2 reinvigorates antitumor immunity and leads to durable tumor regression in mice when combined with osimertinib.DiscussionOur results reveal a new mechanism of EGFR-TKI resistance and suggest a new therapeutic strategy for the treatment of EGFR-mutant tumors.
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- 2023
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3. p16INK4A-deficiency predicts response to combined HER2 and CDK4/6 inhibition in HER2+ breast cancer brain metastases
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Jing Ni, Sheheryar Kabraji, Shaozhen Xie, Yanzhi Wang, Peichen Pan, Xiaofang He, Zongming Liu, Jose Palbo Leone, Henry W. Long, Myles A. Brown, Eric P. Winer, Deborah A. R. Dillon, Nancy U. Lin, and Jean J. Zhao
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Science - Abstract
HER2+ breast cancer often develop brain metastases (BCBMs) that are difficult to treat. Here, the authors show that p16INK4A loss in BCBMs from HER2+ breast tumors results in resistance to the HER2 inhibitor Tucatinib, and that CDK4/6 inhibition can restore sensitivity to this drug.
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- 2022
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4. The role of the PIK3CA gene in the development and aging of the brain
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Shaozhen Xie, Jing Ni, Hanbing Guo, Victor Luu, Yanzhi Wang, Jean J. Zhao, and Thomas M. Roberts
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Medicine ,Science - Abstract
Abstract The CLOVES syndrome is an overgrowth disease arising from mosaic activating somatic mutations in the PIK3CA gene. These mutations occur during fetal development producing malformation and overgrowth of a variety of tissues. It has recently been shown that treatment with low doses of a selective inhibitor of Class I PI3K catalytic subunit p110α, the protein product of the PIK3CA gene, can yield dramatic therapeutic benefits for patients with CLOVES and PROS (a spectrum of PIK3CA-related overgrowth syndromes). To assess the long-term effects of moderate loses of p110α activity, we followed development and growth of mice with heterozygous loss of p110α (Pik3ca +/−) over their entire lifetimes, paying particular attention to effects on the brain. While homozygous deletion of the Pik3ca gene is known to result in early embryonic lethality, these Pik3ca +/− mice displayed a longer lifespan compared to their wild-type littermates. These mice appeared normal, exhibited no obvious behavioral abnormalities, and no body weight changes. However, their brains showed a significant reduction in size and weight. Notably, mice featuring deletion of one allele of Pik3ca only in the brain also showed gradually reduced brain size and weight. Mechanistically, either deletion of p110α or pharmacological inhibition of p110α activity reduced neurosphere size, but not numbers, in vitro, suggesting that p110α activity is critical for neuronal stem cells. The phenotypes observed in our two genetically engineered mouse models suggest that the sustained pharmacological inhibition of the PIK3CA activity in human patients might have both beneficial and harmful effects, and future treatments may need to be deployed in a way to avoid or minimize adverse effects.
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- 2021
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5. A Conditional Dependency on MELK for the Proliferation of Triple-Negative Breast Cancer Cells
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Yubao Wang, Ben B. Li, Jing Li, Thomas M. Roberts, and Jean J. Zhao
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Science - Abstract
Summary: The role of maternal and embryonic leucine zipper kinase (MELK) in cancer cell proliferation has been contentious, with recent studies arriving at disparate conclusions. We investigated the in vitro dependency of cancer cells on MELK under a range of assay conditions. Abrogation of MELK expression has little effect under common culture conditions, in which cells are seeded at high densities and reach confluence in 3–5 days. However, MELK dependency becomes clearly apparent in clonogenic growth assays using either RNAi or CRISPR technologies to modulate MELK expression. This dependency is in sharp contrast to that of essential genes, such as those encoding classic mitotic kinases, but is similar to that of other oncogenes including MYC and KRAS. Our study provides an example demonstrating some of the challenges encountered in cancer target validation, and reveals how subtle, but important, technical variations can ultimately lead to divergent outcomes and conclusions. : Techniques in Genetics; Technical Aspects of Cell Biology; Cancer Subject Areas: Techniques in Genetics, Technical Aspects of Cell Biology, Cancer
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- 2018
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6. CRKL Mediates p110β-Dependent PI3K Signaling in PTEN-Deficient Cancer Cells
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Jing Zhang, Xueliang Gao, Fabienne Schmit, Guillaume Adelmant, Michael J. Eck, Jarrod A. Marto, Jean J. Zhao, and Thomas M. Roberts
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PI3K ,p110β ,CRKL ,Src ,p130Cas ,PTEN ,protein interaction ,cancer ,proliferation ,signaling ,Biology (General) ,QH301-705.5 - Abstract
The p110β isoform of PI3K is preferentially activated in many tumors deficient in the phosphatase and tensin homolog (PTEN). However, the mechanism(s) linking PTEN loss to p110β activation remain(s) mysterious. Here, we identify CRKL as a member of the class of PI3Kβ-interacting proteins. Silencing CRKL expression in PTEN-null human cancer cells leads to a decrease in p110β-dependent PI3K signaling and cell proliferation. In contrast, CRKL depletion does not impair p110α-mediated signaling. Further study showed that CRKL binds to tyrosine-phosphorylated p130Cas in PTEN-null cancer cells. Since Src family kinases are known both to be regulated by PTEN and to phosphorylate and activate p130Cas, we tested and found that Src inhibition cooperated with p110β inhibition to suppress the growth of PTEN-null cells. These data suggest both a potential mechanism linking PTEN loss to p110β activation and the possible benefit of dual inhibition of Src and PI3K for PTEN-null tumors.
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- 2017
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7. PARP Inhibition Elicits STING-Dependent Antitumor Immunity in Brca1-Deficient Ovarian Cancer
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Liya Ding, Hye-Jung Kim, Qiwei Wang, Michael Kearns, Tao Jiang, Carolynn E. Ohlson, Ben B. Li, Shaozhen Xie, Joyce F. Liu, Elizabeth H. Stover, Brooke E. Howitt, Roderick T. Bronson, Suzan Lazo, Thomas M. Roberts, Gordon J. Freeman, Panagiotis A. Konstantinopoulos, Ursula A. Matulonis, and Jean J. Zhao
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Biology (General) ,QH301-705.5 - Abstract
Summary: PARP inhibitors have shown promising clinical activities for patients with BRCA mutations and are changing the landscape of ovarian cancer treatment. However, the therapeutic mechanisms of action for PARP inhibition in the interaction of tumors with the tumor microenvironment and the host immune system remain unclear. We find that PARP inhibition by olaparib triggers robust local and systemic antitumor immunity involving both adaptive and innate immune responses through a STING-dependent antitumor immune response in mice bearing Brca1-deficient ovarian tumors. This effect is further augmented when olaparib is combined with PD-1 blockade. Our findings thus provide a molecular mechanism underlying antitumor activity by PARP inhibition and lay a foundation to improve therapeutic outcome for cancer patients. : Ding et al. show that PARP inhibition in Brca1-deficient tumors elicits strong antitumor immunity involving activation of both innate and adaptive immune responses, a process that is dependent on STING pathway activation. In addition, they show that addition of PD-1 blockade augments the therapeutic efficacy of PARP inhibitor treatment. Keywords: PARP inhibition, BRCA deficiency, STING, immune response, PD-1 blockade, ovarian cancer, targeted therapy, immunotherapy, GEMM
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- 2018
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8. PI3K/AKT Signaling Regulates H3K4 Methylation in Breast Cancer
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Jennifer M. Spangle, Koen M. Dreijerink, Anna C. Groner, Hailing Cheng, Carolynn E. Ohlson, Jaime Reyes, Charles Y. Lin, James Bradner, Jean J. Zhao, Thomas M. Roberts, and Myles Brown
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Biology (General) ,QH301-705.5 - Abstract
Post-translational histone H3 modifications regulate transcriptional competence. The mechanisms by which the epigenome is regulated in response to oncogenic signaling remain unclear. Here we show that H3K4me3 is increased in breast tumors driven by an activated PIK3CA allele and that inhibition of PI3K/AKT signaling reduces promoter-associated H3K4me3 in human breast cancer cells. We show that the H3K4 demethylase KDM5A is an AKT target and that phosphorylation of KDM5A regulates its nuclear localization and promoter occupancy. Supporting a role for KDM5A in mediating PI3K/AKT transcriptional effects, the decreased expression in response to AKT inhibition of a subset of cell-cycle genes associated with poor clinical outcome is blunted by KDM5A silencing. Our data identify a mechanism by which PI3K/AKT signaling modulates the cancer epigenome through controlling H3K4 methylation and suggest that KDM5A subcellular localization and genome occupancy may be pharmacodynamic markers of the activity of PI3K/AKT inhibitors currently in clinical development.
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- 2016
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9. A genetic screen for candidate tumor suppressors identifies REST
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Westbrook, Thomas F., Yumei Leng, Jean J. Zhao, Mandel, Galil, Chin, Lynda, Martin, Eric S., Schlabach, Michael R., Liang, Anthony C., Bin Feng, Roberts, Thomas M., Hannon, Gregory J., DePinho, Ronald A., and Elledge, Stephen J.
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Epithelial cells -- Research ,Cancer -- Genetic aspects ,Cancer -- Research ,Biological sciences - Abstract
An RNAi-based genetic screen for genes that suppress transformation of human mammary epithelial cells is reported. The results implicate REST gene as a human tumor suppressor and provide a novel approach to identifying candidate genes that suppress the development of human cancer.
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- 2005
10. Class IA Phosphatidylinositol 3-Kinase Isoform p110a Mediates Vascular Remodeling.
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Vantler, Marius, Jesus, Joana, Leppänen, Olli, Scherner, Maximilian, Berghausen, Eva Maria, Mustafov, Lenard, Xin Chen, Kramer, Tilmann, Zierden, Mario, Gerhardt, Maximilian, Freyhaus, Henrik ten, Blaschke, Florian, Sterner-Kock, Anja, Baldus, Stephan, Jean J. Zhao, and Rosenkranz, Stephan
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- 2015
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11. STING agonism overcomes STAT3-mediated immunosuppression and adaptive resistance to PARP inhibition in ovarian cancer
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Changli Qian, Gordon J Freeman, Tao Jiang, Ursula Matulonis, Ziying Lin, Hye-Jung Kim, Anniina Färkkilä, Hua Yu, Xin Cheng, Antons Martincuks, Joyce F Liu, Qiwei Wang, Liya Ding, Michael J Kearns, Shaozhen Xie, Inga-Maria Launonen, Thomas M Roberts, and Jean J Zhao
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Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Background Poly (ADP-ribose) polymerase (PARP) inhibition (PARPi) has demonstrated potent therapeutic efficacy in patients with BRCA-mutant ovarian cancer. However, acquired resistance to PARPi remains a major challenge in the clinic.Methods PARPi-resistant ovarian cancer mouse models were generated by long-term treatment of olaparib in syngeneic Brca1-deficient ovarian tumors. Signal transducer and activator of transcription 3 (STAT3)-mediated immunosuppression was investigated in vitro by co-culture experiments and in vivo by analysis of immune cells in the tumor microenvironment (TME) of human and mouse PARPi-resistant tumors. Whole genome transcriptome analysis was performed to assess the antitumor immunomodulatory effect of STING (stimulator of interferon genes) agonists on myeloid cells in the TME of PARPi-resistant ovarian tumors. A STING agonist was used to overcome STAT3-mediated immunosuppression and acquired PARPi resistance in syngeneic and patient-derived xenografts models of ovarian cancer.Results In this study, we uncover an adaptive resistance mechanism to PARP inhibition mediated by tumor-associated macrophages (TAMs) in the TME. Markedly increased populations of protumor macrophages are found in BRCA-deficient ovarian tumors that rendered resistance to PARPi in both murine models and patients. Mechanistically, PARP inhibition elevates the STAT3 signaling pathway in tumor cells, which in turn promotes protumor polarization of TAMs. STAT3 ablation in tumor cells mitigates polarization of protumor macrophages and increases tumor-infiltrating T cells on PARP inhibition. These findings are corroborated in patient-derived, PARPi-resistant BRCA1-mutant ovarian tumors. Importantly, STING agonists reshape the immunosuppressive TME by reprogramming myeloid cells and overcome the TME-dependent adaptive resistance to PARPi in ovarian cancer. This effect is further enhanced by addition of the programmed cell death protein-1 blockade.Conclusions We elucidate an adaptive immunosuppression mechanism rendering resistance to PARPi in BRCA1-mutant ovarian tumors. This is mediated by enrichment of protumor TAMs propelled by PARPi-induced STAT3 activation in tumor cells. We also provide a new strategy to reshape the immunosuppressive TME with STING agonists and overcome PARPi resistance in ovarian cancer.
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- 2023
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12. An alternative splicing switch in FLNB promotes the mesenchymal cell state in human breast cancer
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Ji Li, Peter S Choi, Christine L Chaffer, Katherine Labella, Justin H Hwang, Andrew O Giacomelli, Jong Wook Kim, Nina Ilic, John G Doench, Seav Huong Ly, Chao Dai, Kimberly Hagel, Andrew L Hong, Ole Gjoerup, Shom Goel, Jennifer Y Ge, David E Root, Jean J Zhao, Angela N Brooks, Robert A Weinberg, and William C Hahn
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alternative splicing ,QKI ,RBFOX1 ,FLNB ,EMT ,basal-like breast cancer ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Alternative splicing of mRNA precursors represents a key gene expression regulatory step and permits the generation of distinct protein products with diverse functions. In a genome-scale expression screen for inducers of the epithelial-to-mesenchymal transition (EMT), we found a striking enrichment of RNA-binding proteins. We validated that QKI and RBFOX1 were necessary and sufficient to induce an intermediate mesenchymal cell state and increased tumorigenicity. Using RNA-seq and eCLIP analysis, we found that QKI and RBFOX1 coordinately regulated the splicing and function of the actin-binding protein FLNB, which plays a causal role in the regulation of EMT. Specifically, the skipping of FLNB exon 30 induced EMT by releasing the FOXC1 transcription factor. Moreover, skipping of FLNB exon 30 is strongly associated with EMT gene signatures in basal-like breast cancer patient samples. These observations identify a specific dysregulation of splicing, which regulates tumor cell plasticity and is frequently observed in human cancer.
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- 2018
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13. Correction: MELK is an oncogenic kinase essential for mitotic progression in basal-like breast cancer cells
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Yubao Wang, Young-Mi Li, Lukas Baitsch, Alan Huang, Yi Xiang, Haoxuan Tong, Ana Lako, Thanh Von, Christine Choi, Elgene Lim, Junxia Min, Li Li, Frank Stegmeier, Robert Schlegel, Michael J Eck, Nathanael S Gray, Timothy J Mitchison, and Jean J Zhao
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Medicine ,Science ,Biology (General) ,QH301-705.5 - Published
- 2018
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14. Pik3ca is required for mouse uterine gland development and pregnancy.
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Hye Jin Chang, Hee Sung Shin, Tae Hoon Kim, Jung-Yoon Yoo, Hanna E Teasley, Jean J Zhao, Un-Hwan Ha, and Jae-Wook Jeong
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Medicine ,Science - Abstract
The PI3K/AKT signaling pathway plays a critical role in the maintenance of equilibrium between cell survival and apoptosis. The Pik3ca gene is mutated in a range of human cancers. It has been found to be oncogenic, and mutations lead to constitutive activation of the PI3K/AKT pathway. The expression patterns of PIK3CA proteins in the uterus of mice during early pregnancy indicate that it may play a role in the regulation of glandular epithelial cells, which is required to support uterine receptivity. To further investigate the role of Pik3ca in uterine function, Pik3ca was conditionally ablated only in the PGR-positive cells (Pgrcre/+Pik3caf/f; Pik3cad/d). A defect of uterine gland development and decidualization led to subfertility observed in Pik3cad/d mice. Pik3cad/d mice showed significantly decreased uterine weight compared to Pik3caf/f mice. Interestingly, a significant decrease of gland numbers were detected in Pik3cad/d mice compared to control mice. In addition, we found a decrease of Foxa2 expression, which is a known uterine gland marker in Pik3cad/d mice. Furthermore, the excessive proliferation of endometrial epithelial cells was observed in Pik3cad/d mice. Our studies suggest that Pik3ca has a critical role in uterine gland development and female fertility.
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- 2018
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15. MELK is not necessary for the proliferation of basal-like breast cancer cells
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Hai-Tsang Huang, Hyuk-Soo Seo, Tinghu Zhang, Yubao Wang, Baishan Jiang, Qing Li, Dennis L Buckley, Behnam Nabet, Justin M Roberts, Joshiawa Paulk, Shiva Dastjerdi, Georg E Winter, Hilary McLauchlan, Jennifer Moran, James E Bradner, Michael J Eck, Sirano Dhe-Paganon, Jean J Zhao, and Nathanael S Gray
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MELK ,basal-like breast cancer ,OTSSP167 ,HTH-01-091 ,target validation ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Thorough preclinical target validation is essential for the success of drug discovery efforts. In this study, we combined chemical and genetic perturbants, including the development of a novel selective maternal embryonic leucine zipper kinase (MELK) inhibitor HTH-01-091, CRISPR/Cas9-mediated MELK knockout, a novel chemical-induced protein degradation strategy, RNA interference and CRISPR interference to validate MELK as a therapeutic target in basal-like breast cancers (BBC). In common culture conditions, we found that small molecule inhibition, genetic deletion, or acute depletion of MELK did not significantly affect cellular growth. This discrepancy to previous findings illuminated selectivity issues of the widely used MELK inhibitor OTSSP167, and potential off-target effects of MELK-targeting short hairpins. The different genetic and chemical tools developed here allow for the identification and validation of any causal roles MELK may play in cancer biology, which will be required to guide future MELK drug discovery efforts. Furthermore, our study provides a general framework for preclinical target validation.
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- 2017
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16. Rac1-mediated membrane raft localization of PI3K/p110β is required for its activation by GPCRs or PTEN loss
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Onur Cizmecioglu, Jing Ni, Shaozhen Xie, Jean J Zhao, and Thomas M Roberts
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PI3K signaling ,Rac1 ,PTEN loss ,GPCR signaling ,membrane microdomains ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
We aimed to understand how spatial compartmentalization in the plasma membrane might contribute to the functions of the ubiquitous class IA phosphoinositide 3-kinase (PI3K) isoforms, p110α and p110β. We found that p110β localizes to membrane rafts in a Rac1-dependent manner. This localization potentiates Akt activation by G-protein-coupled receptors (GPCRs). Thus genetic targeting of a Rac1 binding-deficient allele of p110β to rafts alleviated the requirement for p110β-Rac1 association for GPCR signaling, cell growth and migration. In contrast, p110α, which does not play a physiological role in GPCR signaling, is found to reside in nonraft regions of the plasma membrane. Raft targeting of p110α allowed its EGFR-mediated activation by GPCRs. Notably, p110β dependent, PTEN null tumor cells critically rely upon raft-associated PI3K activity. Collectively, our findings provide a mechanistic account of how membrane raft localization regulates differential activation of distinct PI3K isoforms and offer insight into why PTEN-deficient cancers depend on p110β.
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- 2016
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17. Mouse models of human PIK3CA-related brain overgrowth have acutely treatable epilepsy
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Achira Roy, Jonathan Skibo, Franck Kalume, Jing Ni, Sherri Rankin, Yiling Lu, William B Dobyns, Gordon B Mills, Jean J Zhao, Suzanne J Baker, and Kathleen J Millen
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PI3K ,megalencephaly ,mouse models ,epilepsy ,BKM120 ,cortical dysplasia ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Mutations in the catalytic subunit of phosphoinositide 3-kinase (PIK3CA) and other PI3K-AKT pathway components have been associated with cancer and a wide spectrum of brain and body overgrowth. In the brain, the phenotypic spectrum of PIK3CA-related segmental overgrowth includes bilateral dysplastic megalencephaly, hemimegalencephaly and focal cortical dysplasia, the most common cause of intractable pediatric epilepsy. We generated mouse models expressing the most common activating Pik3ca mutations (H1047R and E545K) in developing neural progenitors. These accurately recapitulate all the key human pathological features including brain enlargement, cortical malformation, hydrocephalus and epilepsy, with phenotypic severity dependent on the mutant allele and its time of activation. Underlying mechanisms include increased proliferation, cell size and altered white matter. Notably, we demonstrate that acute 1 hr-suppression of PI3K signaling despite the ongoing presence of dysplasia has dramatic anti-epileptic benefit. Thus PI3K inhibitors offer a promising new avenue for effective anti-epileptic therapy for intractable pediatric epilepsy patients.
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- 2015
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18. MELK is an oncogenic kinase essential for mitotic progression in basal-like breast cancer cells
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Yubao Wang, Young-Mi Lee, Lukas Baitsch, Alan Huang, Yi Xiang, Haoxuan Tong, Ana Lako, Thanh Von, Christine Choi, Elgene Lim, Junxia Min, Li Li, Frank Stegmeier, Robert Schlegel, Michael J Eck, Nathanael S Gray, Timothy J Mitchison, and Jean J Zhao
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MELK ,basal-like breast cancer ,triple-negative breast cancer ,FoxM1 ,targeted therapy ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
Despite marked advances in breast cancer therapy, basal-like breast cancer (BBC), an aggressive subtype of breast cancer usually lacking estrogen and progesterone receptors, remains difficult to treat. In this study, we report the identification of MELK as a novel oncogenic kinase from an in vivo tumorigenesis screen using a kinome-wide open reading frames (ORFs) library. Analysis of clinical data reveals a high level of MELK overexpression in BBC, a feature that is largely dependent on FoxM1, a master mitotic transcription factor that is also found to be highly overexpressed in BBC. Ablation of MELK selectively impairs proliferation of basal-like, but not luminal breast cancer cells both in vitro and in vivo. Mechanistically, depletion of MELK in BBC cells induces caspase-dependent cell death, preceded by defective mitosis. Finally, we find that Melk is not required for mouse development and physiology. Together, these data indicate that MELK is a normally non-essential kinase, but is critical for BBC and thus represents a promising selective therapeutic target for the most aggressive subtype of breast cancer.
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- 2014
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19. Therapeutic implications of GIPC1 silencing in cancer.
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Thomas W Chittenden, Jane Pak, Renee Rubio, Hailing Cheng, Kristina Holton, Niall Prendergast, Vladimir Glinskii, Yi Cai, Aedin Culhane, Stefan Bentink, Mathew Schwede, Jessica C Mar, Eleanor A Howe, Martin Aryee, Razvan Sultana, Anthony A Lanahan, Jennifer M Taylor, Chris Holmes, William C Hahn, Jean J Zhao, J Dirk Iglehart, and John Quackenbush
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Medicine ,Science - Abstract
GIPC1 is a cytoplasmic scaffold protein that interacts with numerous receptor signaling complexes, and emerging evidence suggests that it plays a role in tumorigenesis. GIPC1 is highly expressed in a number of human malignancies, including breast, ovarian, gastric, and pancreatic cancers. Suppression of GIPC1 in human pancreatic cancer cells inhibits in vivo tumor growth in immunodeficient mice. To better understand GIPC1 function, we suppressed its expression in human breast and colorectal cancer cell lines and human mammary epithelial cells (HMECs) and assayed both gene expression and cellular phenotype. Suppression of GIPC1 promotes apoptosis in MCF-7, MDA-MD231, SKBR-3, SW480, and SW620 cells and impairs anchorage-independent colony formation of HMECs. These observations indicate GIPC1 plays an essential role in oncogenic transformation, and its expression is necessary for the survival of human breast and colorectal cancer cells. Additionally, a GIPC1 knock-down gene signature was used to interrogate publically available breast and ovarian cancer microarray datasets. This GIPC1 signature statistically correlates with a number of breast and ovarian cancer phenotypes and clinical outcomes, including patient survival. Taken together, these data indicate that GIPC1 inhibition may represent a new target for therapeutic development for the treatment of human cancers.
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- 2010
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20. Breakage fusion bridge cycles drive high oncogene copy number, but not intratumoral genetic heterogeneity or rapid cancer genome change.
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Dehkordi SR, Wong IT, Ni J, Luebeck J, Zhu K, Prasad G, Krockenberger L, Xu G, Chowdhury B, Rajkumar U, Caplin A, Muliaditan D, Coruh C, Jin Q, Turner K, Teo SX, Pang AWC, Alexandrov LB, Chua CEL, Furnari FB, Paulson TG, Law JA, Chang HY, Yue F, DasGupta R, Zhao J, Mischel PS, and Bafna V
- Abstract
Oncogene amplification is a major driver of cancer pathogenesis. Breakage fusion bridge (BFB) cycles, like extrachromosomal DNA (ecDNA), can lead to high copy numbers of oncogenes, but their impact on intratumoral heterogeneity, treatment response, and patient survival are not well understood due to difficulty in detecting them by DNA sequencing. We describe a novel algorithm that detects and reconstructs BFB amplifications using optical genome maps (OGMs), called OM2BFB. OM2BFB showed high precision (>93%) and recall (92%) in detecting BFB amplifications in cancer cell lines, PDX models and primary tumors. OM-based comparisons demonstrated that short-read BFB detection using our AmpliconSuite (AS) toolkit also achieved high precision, albeit with reduced sensitivity. We detected 371 BFB events using whole genome sequences from 2,557 primary tumors and cancer lines. BFB amplifications were preferentially found in cervical, head and neck, lung, and esophageal cancers, but rarely in brain cancers. BFB amplified genes show lower variance of gene expression, with fewer options for regulatory rewiring relative to ecDNA amplified genes. BFB positive (BFB (+)) tumors showed reduced heterogeneity of amplicon structures, and delayed onset of resistance, relative to ecDNA(+) tumors. EcDNA and BFB amplifications represent contrasting mechanisms to increase the copy numbers of oncogene with markedly different characteristics that suggest different routes for intervention.
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- 2023
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21. Lacrimal gland budding requires PI3K-dependent suppression of EGF signaling.
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Wang Q, Tao C, Hannan A, Yoon S, Min X, Peregrin J, Qu X, Li H, Yu H, Zhao J, and Zhang X
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The patterning of epithelial buds is determined by the underlying signaling network. Here, we study the cross-talk between phosphoinositide 3-kinase (PI3K) and Ras signaling during lacrimal gland budding morphogenesis. Our results show that PI3K is activated by both the p85-mediated insulin-like growth factor (IGF) and Ras-mediated fibroblast growth factor (FGF) signaling. On the other hand, PI3K also promotes extracellular signal-regulated kinase (ERK) signaling via a direct interaction with Ras. Both PI3K and ERK are upstream regulators of mammalian target of rapamycin (mTOR), and, together, they prevent expansion of epidermal growth factor (EGF) receptor expression from the lacrimal gland stalk to the bud region. We further show that this suppression of EGF signaling is necessary for induction of lacrimal gland buds. These results reveal that the interplay between PI3K, mitogen-activated protein kinase, and mTOR mediates the cross-talk among FGF, IGF, and EGF signaling in support of lacrimal gland development., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)
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- 2021
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22. A sequential methodology for the rapid identification and characterization of breast cancer-associated functional SNPs.
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Zhao Y, Wu D, Jiang D, Zhang X, Wu T, Cui J, Qian M, Zhao J, Oesterreich S, Sun W, Finkel T, and Li G
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- Adaptor Proteins, Signal Transducing genetics, Blotting, Western, Breast Neoplasms pathology, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Humans, Linkage Disequilibrium, MAP Kinase Kinase Kinase 1 genetics, MCF-7 Cells, Mass Spectrometry methods, Receptor, Fibroblast Growth Factor, Type 2 genetics, Regulatory Sequences, Nucleic Acid genetics, Breast Neoplasms genetics, Genetic Predisposition to Disease genetics, Genome-Wide Association Study methods, Polymorphism, Single Nucleotide, Sequence Analysis, DNA methods
- Abstract
GWAS cannot identify functional SNPs (fSNP) from disease-associated SNPs in linkage disequilibrium (LD). Here, we report developing three sequential methodologies including Reel-seq (Regulatory element-sequencing) to identify fSNPs in a high-throughput fashion, SDCP-MS (SNP-specific DNA competition pulldown-mass spectrometry) to identify fSNP-bound proteins and AIDP-Wb (allele-imbalanced DNA pulldown-Western blot) to detect allele-specific protein:fSNP binding. We first apply Reel-seq to screen a library containing 4316 breast cancer-associated SNPs and identify 521 candidate fSNPs. As proof of principle, we verify candidate fSNPs on three well-characterized loci: FGFR2, MAP3K1 and BABAM1. Next, using SDCP-MS and AIDP-Wb, we rapidly identify multiple regulatory factors that specifically bind in an allele-imbalanced manner to the fSNPs on the FGFR2 locus. We finally demonstrate that the factors identified by SDCP-MS can regulate risk gene expression. These data suggest that the sequential application of Reel-seq, SDCP-MS, and AIDP-Wb can greatly help to translate large sets of GWAS data into biologically relevant information.
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- 2020
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23. Dual HDAC and PI3K Inhibition Abrogates NFκB- and FOXM1-Mediated DNA Damage Response to Radiosensitize Pediatric High-Grade Gliomas.
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Pal S, Kozono D, Yang X, Fendler W, Fitts W, Ni J, Alberta JA, Zhao J, Liu KX, Bian J, Truffaux N, Weiss WA, Resnick AC, Bandopadhayay P, Ligon KL, DuBois SG, Mueller S, Chowdhury D, and Haas-Kogan DA
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- Animals, Brain Neoplasms drug therapy, Brain Neoplasms metabolism, Cell Line, Tumor, Child, DNA Repair drug effects, Glioma metabolism, Homologous Recombination drug effects, Humans, Mice, Mice, Nude, Morpholines pharmacology, Pyrimidines pharmacology, Radiation-Sensitizing Agents pharmacology, DNA Damage drug effects, Forkhead Box Protein M1 metabolism, Glioma drug therapy, Histone Deacetylase Inhibitors pharmacology, Histone Deacetylases metabolism, NF-kappa B metabolism, Phosphoinositide-3 Kinase Inhibitors
- Abstract
Aberrant chromatin remodeling and activation of the PI3K pathway have been identified as important mediators of pediatric high-grade glioma (pHGG) and diffuse intrinsic pontine glioma (DIPG) pathogenesis. As inhibition of these pathways are promising therapeutic avenues and radiation is the only modality to prolong survival of patients with DIPG, we sought to explore radiosensitizing functions of such inhibition and to explore mechanisms of action of such agents. Here, we demonstrate that combined treatment with radiotherapy and CUDC-907, a novel first-in-class dual inhibitor of histone deacetylases (HDAC) and PI3K, evokes a potent cytotoxic response in pHGG and DIPG models. CUDC-907 modulated DNA damage response by inhibiting radiation-induced DNA repair pathways including homologous recombination and nonhomologous end joining. The radiosensitizing effects of CUDC-907 were mediated by decreased NFκB/Forkhead box M1 (FOXM1) recruitment to promoters of genes involved in the DNA damage response; exogenous expression of NFκB/FOXM1 protected from CUDC-907-induced cytotoxicity. Together, these findings reveal CUDC-907 as a novel radiosensitizer with potent antitumor activity in pHGG and DIPG and provide a preclinical rationale for the combination of CUDC-907 with radiotherapy as a novel therapeutic strategy against pHGG and DIPG. More globally, we have identified NFκB and FOXM1 and their downstream transcriptional elements as critical targets for new treatments for pHGG and DIPG. Significance: These findings describe the radiosensitizing effect of a novel agent in pediatric high-grade gliomas, addressing a critical unmet need of increasing the radiation sensitivity of these highly aggressive tumors. Cancer Res; 78(14); 4007-21. ©2018 AACR ., (©2018 American Association for Cancer Research.)
- Published
- 2018
- Full Text
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24. Isoform-Selective Phosphatidylinositol 3-Kinase Inhibition in Cancer.
- Author
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Bergholz JS, Roberts TM, and Zhao JJ
- Subjects
- Class I Phosphatidylinositol 3-Kinases, Humans, Protein Isoforms, Thiazoles, Neoplasms, Phosphatidylinositol 3-Kinase
- Published
- 2018
- Full Text
- View/download PDF
25. Allele-Specific Chromatin Recruitment and Therapeutic Vulnerabilities of ESR1 Activating Mutations.
- Author
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Jeselsohn R, Bergholz JS, Pun M, Cornwell M, Liu W, Nardone A, Xiao T, Li W, Qiu X, Buchwalter G, Feiglin A, Abell-Hart K, Fei T, Rao P, Long H, Kwiatkowski N, Zhang T, Gray N, Melchers D, Houtman R, Liu XS, Cohen O, Wagle N, Winer EP, Zhao J, and Brown M
- Subjects
- Animals, Antineoplastic Agents, Hormonal pharmacology, Breast Neoplasms genetics, Breast Neoplasms pathology, Drug Resistance, Neoplasm genetics, Humans, Mice, Transgenic, Alleles, Chromatin metabolism, Estrogen Receptor alpha genetics, Mutation genetics
- Abstract
Estrogen receptor α (ER) ligand-binding domain (LBD) mutations are found in a substantial number of endocrine treatment-resistant metastatic ER-positive (ER
+ ) breast cancers. We investigated the chromatin recruitment, transcriptional network, and genetic vulnerabilities in breast cancer models harboring the clinically relevant ER mutations. These mutants exhibit both ligand-independent functions that mimic estradiol-bound wild-type ER as well as allele-specific neomorphic properties that promote a pro-metastatic phenotype. Analysis of the genome-wide ER binding sites identified mutant ER unique recruitment mediating the allele-specific transcriptional program. Genetic screens identified genes that are essential for the ligand-independent growth driven by the mutants. These studies provide insights into the mechanism of endocrine therapy resistance engendered by ER mutations and potential therapeutic targets., (Copyright © 2018 Elsevier Inc. All rights reserved.)- Published
- 2018
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- View/download PDF
26. Quantifying survival in patients with Proteus syndrome.
- Author
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Sapp JC, Hu L, Zhao J, Gruber A, Schwartz B, Ferrari D, and Biesecker LG
- Subjects
- Adolescent, Child, Child, Preschool, Female, Humans, Infant, Infant, Newborn, Kaplan-Meier Estimate, Male, Proteus Syndrome epidemiology, Young Adult, Proteus Syndrome mortality
- Abstract
PurposeProteus syndrome is a rare mosaic overgrowth disorder that is associated with severe complications. While anecdotal data have suggested that the life span of affected patients is reduced, this has not been measured. Mortality data on rare diseases is critical for assessing treatments and other interventions.MethodsTo address this we used the clinical research records of 64 patients in a longitudinal natural history cohort at the National Institutes of Health to ascertain the data in an organized manner and estimate survival using a Kaplan-Meier approach.ResultsThe median age of diagnosis was 19 months. Based on this analysis, there was 25% probability of death by 22 years of age. Ten of the 11 patients who died were younger than 22 years of age, and there was only a single death after this age.ConclusionThese data quantify the risk of premature death in Proteus syndrome, which can be used to support interventions and trials. Although the risk of death is substantial, the fact that only one patient died after 22 years of age supports anecdotal evidence that the disease process moderates after the end of adolescence. Interventions to reduce mortality should be targeted to the pediatric age range.
- Published
- 2017
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27. A cell-based quantitative high-throughput image screening identified novel autophagy modulators.
- Author
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Li Y, McGreal S, Zhao J, Huang R, Zhou Y, Zhong H, Xia M, and Ding WX
- Subjects
- A549 Cells, Animals, Antineoplastic Agents pharmacology, Chlorpromazine pharmacology, Dopamine Antagonists pharmacology, Dose-Response Relationship, Drug, Fibroblasts metabolism, Fibroblasts pathology, Fluphenazine pharmacology, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, HCT116 Cells, Humans, Indans pharmacology, Methylamines pharmacology, Mice, Microtubule-Associated Proteins genetics, Microtubule-Associated Proteins metabolism, Neoplasms metabolism, Neoplasms pathology, Protein Kinase Inhibitors pharmacology, Receptors, Dopamine drug effects, Receptors, Dopamine metabolism, Recombinant Fusion Proteins metabolism, Signal Transduction drug effects, TOR Serine-Threonine Kinases antagonists & inhibitors, TOR Serine-Threonine Kinases metabolism, Transfection, Autophagy drug effects, Biological Assay, Drug Discovery methods, Fibroblasts drug effects, High-Throughput Screening Assays, Neoplasms drug therapy
- Abstract
Macroautophagy is a major cellular degradation pathway for long-lived proteins and cellular organelles to maintain cellular homeostasis. Reduced autophagy has been implicated in neurodegenerative diseases, metabolic syndrome, and tumorigenesis. In contrast, increased autophagy has been shown to protect against tissue injury and aging. Here we employed a cell-based quantitative high-throughput image screening (qHTS) for autophagy modulators using mouse embryonic fibroblasts (MEFs) that are stably expressing GFP-LC3. The library of pharmacologically active compounds (LOPAC) was used to screen for the autophagy modulators in compounds alone or in combination with the lysosome inhibitor chloroquine (CQ). The GFP-LC3 puncta were then quantified to measure autophagic flux. The primary screening revealed 173 compounds with efficacy more than 40%. These compounds were cherry-picked and re-tested at multiple different concentrations using the same assay. A number of novel autophagy inducers, inhibitors, and modulators with dual-effects on autophagy were identified from the cherry-pick screening. Interestingly, we found a group of compounds that induce autophagy are related to dopamine receptors and are commonly used as clinical psychiatric drugs. Among them, indatraline hydrochloride (IND), a dopamine inhibitor, and chlorpromazine hydrochloride (CPZ) and fluphenazine dihydrochloride (FPZ), two dopamine receptor antagonists, were further evaluated. We found that FPZ-induced autophagy through mTOR inhibition but IND and CPZ induced autophagy in an mTOR-independent manner. Our data suggest that image-based autophagic flux qHTS can efficiently identify autophagy inducers and inhibitors., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
28. High-Throughput Phenotypic Screening of Human Astrocytes to Identify Compounds That Protect Against Oxidative Stress.
- Author
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Thorne N, Malik N, Shah S, Zhao J, Class B, Aguisanda F, Southall N, Xia M, McKew JC, Rao M, and Zheng W
- Subjects
- Antioxidant Response Elements drug effects, Astrocytes metabolism, Cytoprotection, Dose-Response Relationship, Drug, Embryonic Stem Cells metabolism, Gene Expression Regulation, Developmental, Hep G2 Cells, Humans, Hydrogen Peroxide toxicity, NF-E2-Related Factor 2 genetics, NF-E2-Related Factor 2 metabolism, Oxidants pharmacology, Phenotype, Reproducibility of Results, Small Molecule Libraries, Antioxidants pharmacology, Astrocytes drug effects, Drug Discovery methods, Embryonic Stem Cells drug effects, High-Throughput Screening Assays methods, Neurogenesis, Oxidative Stress drug effects
- Abstract
Unlabelled: Astrocytes are the predominant cell type in the nervous system and play a significant role in maintaining neuronal health and homeostasis. Recently, astrocyte dysfunction has been implicated in the pathogenesis of many neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and amyotrophic lateral sclerosis. Astrocytes are thus an attractive new target for drug discovery for neurological disorders. Using astrocytes differentiated from human embryonic stem cells, we have developed an assay to identify compounds that protect against oxidative stress, a condition associated with many neurodegenerative diseases. This phenotypic oxidative stress assay has been optimized for high-throughput screening in a 1,536-well plate format. From a screen of approximately 4,100 bioactive tool compounds and approved drugs, we identified a set of 22 that acutely protect human astrocytes from the consequences of hydrogen peroxide-induced oxidative stress. Nine of these compounds were also found to be protective of induced pluripotent stem cell-differentiated astrocytes in a related assay. These compounds are thought to confer protection through hormesis, activating stress-response pathways and preconditioning astrocytes to handle subsequent exposure to hydrogen peroxide. In fact, four of these compounds were found to activate the antioxidant response element/nuclear factor-E2-related factor 2 pathway, a protective pathway induced by toxic insults. Our results demonstrate the relevancy and utility of using astrocytes differentiated from human stem cells as a disease model for drug discovery and development., Significance: Astrocytes play a key role in neurological diseases. Drug discovery efforts that target astrocytes can identify novel therapeutics. Human astrocytes are difficult to obtain and thus are challenging to use for high-throughput screening, which requires large numbers of cells. Using human embryonic stem cell-derived astrocytes and an optimized astrocyte differentiation protocol, it was possible to screen approximately 4,100 compounds in titration to identify 22 that are cytoprotective of astrocytes. This study is the largest-scale high-throughput screen conducted using human astrocytes, with a total of 17,536 data points collected in the primary screen. The results demonstrate the relevancy and utility of using astrocytes differentiated from human stem cells as a disease model for drug discovery and development., (©AlphaMed Press.)
- Published
- 2016
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29. PI3K in the ventromedial hypothalamic nucleus mediates estrogenic actions on energy expenditure in female mice.
- Author
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Saito K, He Y, Yang Y, Zhu L, Wang C, Xu P, Hinton AO, Yan X, Zhao J, Fukuda M, Tong Q, Clegg DJ, and Xu Y
- Subjects
- Animals, Body Weight, Disease Models, Animal, Energy Metabolism, Estrogens pharmacology, Female, Male, Mice, Obesity chemically induced, Sex Characteristics, Estrogen Receptor alpha metabolism, Estrogens metabolism, Obesity metabolism, Phosphatidylinositol 3-Kinases metabolism, Ventromedial Hypothalamic Nucleus metabolism
- Abstract
Estrogens act in the ventromedial hypothalamic nucleus (VMH) to regulate body weight homeostasis. However, the molecular mechanisms underlying these estrogenic effects are unknown. We show that activation of estrogen receptor-α (ERα) stimulates neural firing of VMH neurons expressing ERα, and these effects are blocked with intracellular application of a pharmacological inhibitor of the phosphatidyl inositol 3-kinase (PI3K). Further, we demonstrated that mice with genetic inhibition of PI3K activity in VMH neurons showed a sexual dimorphic obese phenotype, with only female mutants being affected. In addition, inhibition of VMH PI3K activity blocked effects of 17β-estradiol to stimulate energy expenditure, but did not affect estrogen-induced anorexia. Collectively, our results indicate that PI3K activity in VMH neurons plays a physiologically relevant role in mediating estrogenic actions on energy expenditure in females.
- Published
- 2016
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30. The ERα-PI3K Cascade in Proopiomelanocortin Progenitor Neurons Regulates Feeding and Glucose Balance in Female Mice.
- Author
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Zhu L, Xu P, Cao X, Yang Y, Hinton AO Jr, Xia Y, Saito K, Yan X, Zou F, Ding H, Wang C, Yan C, Saha P, Khan SA, Zhao J, Fukuda M, Tong Q, Clegg DJ, Chan L, and Xu Y
- Subjects
- Animals, Eating drug effects, Estrogen Receptor alpha agonists, Estrogen Receptor alpha metabolism, Feeding Behavior physiology, Female, Glucose metabolism, Homeostasis, Mice, Mice, Knockout, Neural Stem Cells drug effects, Neurons drug effects, Phenols pharmacology, Phosphatidylinositol 3-Kinase drug effects, Pyrazoles pharmacology, Signal Transduction, Blood Glucose metabolism, Eating genetics, Estrogen Receptor alpha genetics, Estrogens metabolism, Insulin Resistance, Neural Stem Cells metabolism, Neurons metabolism, Phosphatidylinositol 3-Kinase genetics, Pro-Opiomelanocortin metabolism
- Abstract
Estrogens act upon estrogen receptor (ER)α to inhibit feeding and improve glucose homeostasis in female animals. However, the intracellular signals that mediate these estrogenic actions remain unknown. Here, we report that anorexigenic effects of estrogens are blunted in female mice that lack ERα specifically in proopiomelanocortin (POMC) progenitor neurons. These mutant mice also develop insulin resistance and are insensitive to the glucose-regulatory effects of estrogens. Moreover, we showed that propyl pyrazole triol (an ERα agonist) stimulates the phosphatidyl inositol 3-kinase (PI3K) pathway specifically in POMC progenitor neurons, and that blockade of PI3K attenuates propyl pyrazole triol-induced activation of POMC neurons. Finally, we show that effects of estrogens to inhibit food intake and to improve insulin sensitivity are significantly attenuated in female mice with PI3K genetically inhibited in POMC progenitor neurons. Together, our results indicate that an ERα-PI3K cascade in POMC progenitor neurons mediates estrogenic actions to suppress food intake and improve insulin sensitivity.
- Published
- 2015
- Full Text
- View/download PDF
31. Measurement of PIP3 levels reveals an unexpected role for p110β in early adaptive responses to p110α-specific inhibitors in luminal breast cancer.
- Author
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Costa C, Ebi H, Martini M, Beausoleil SA, Faber AC, Jakubik CT, Huang A, Wang Y, Nishtala M, Hall B, Rikova K, Zhao J, Hirsch E, Benes CH, and Engelman JA
- Subjects
- Animals, Breast Neoplasms metabolism, Breast Neoplasms pathology, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases antagonists & inhibitors, Female, Gene Expression Regulation, Neoplastic drug effects, Humans, MCF-7 Cells, Mice, Mice, Nude, Neoplasm Transplantation, Receptor, ErbB-2 genetics, Signal Transduction drug effects, Antineoplastic Combined Chemotherapy Protocols pharmacology, Breast Neoplasms drug therapy, Class I Phosphatidylinositol 3-Kinases metabolism, Pyrimidinones pharmacology, Thiazoles pharmacology, ortho-Aminobenzoates pharmacology
- Abstract
BYL719, which selectively inhibits the alpha isoform of the phosphatidylinositol 3-kinase (PI3K) catalytic subunit (p110a), is currently in clinical trials for the treatment of solid tumors, especially luminal breast cancers with PIK3CA mutations and/or HER2 amplification. This study reveals that, even among these sensitive cancers, the initial efficacy of p110α inhibition is mitigated by rapid re-accumulation of the PI3K product PIP3 produced by the p110β isoform. Importantly, the reactivation of PI3K mediated by p110β does not invariably restore AKT phosphorylation, demonstrating the limitations of using phospho-AKT as a surrogate to measure PI3K activation. Consistently, we show that the addition of the p110β inhibitor to BYL719 prevents the PIP3 rebound and induces greater antitumor efficacy in HER2-amplified and PIK3CA mutant cancers., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
32. Coordinate activation of Shh and PI3K signaling in PTEN-deficient glioblastoma: new therapeutic opportunities.
- Author
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Filbin MG, Dabral SK, Pazyra-Murphy MF, Ramkissoon S, Kung AL, Pak E, Chung J, Theisen MA, Sun Y, Franchetti Y, Sun Y, Shulman DS, Redjal N, Tabak B, Beroukhim R, Wang Q, Zhao J, Dorsch M, Buonamici S, Ligon KL, Kelleher JF, and Segal RA
- Subjects
- Aminopyridines administration & dosage, Animals, Biphenyl Compounds administration & dosage, Brain Neoplasms genetics, Cell Line, Tumor, Enzyme Inhibitors administration & dosage, Glioblastoma genetics, Hedgehog Proteins antagonists & inhibitors, Humans, Mice, Mice, Nude, Morpholines administration & dosage, PTEN Phosphohydrolase genetics, Phosphoinositide-3 Kinase Inhibitors, Pyridines administration & dosage, Ribosomal Protein S6 Kinases antagonists & inhibitors, Signal Transduction drug effects, Xenograft Model Antitumor Assays, Brain Neoplasms drug therapy, Brain Neoplasms metabolism, Glioblastoma drug therapy, Glioblastoma metabolism, Hedgehog Proteins metabolism, PTEN Phosphohydrolase deficiency, Phosphatidylinositol 3-Kinases metabolism
- Abstract
In glioblastoma, phosphatidylinositol 3-kinase (PI3K) signaling is frequently activated by loss of the tumor suppressor phosphatase and tensin homolog (PTEN). However, it is not known whether inhibiting PI3K represents a selective and effective approach for treatment. We interrogated large databases and found that sonic hedgehog (SHH) signaling is activated in PTEN-deficient glioblastoma. We demonstrate that the SHH and PI3K pathways synergize to promote tumor growth and viability in human PTEN-deficient glioblastomas. A combination of PI3K and SHH signaling inhibitors not only suppressed the activation of both pathways but also abrogated S6 kinase (S6K) signaling. Accordingly, targeting both pathways simultaneously resulted in mitotic catastrophe and tumor apoptosis and markedly reduced the growth of PTEN-deficient glioblastomas in vitro and in vivo. The drugs tested here appear to be safe in humans; therefore, this combination may provide a new targeted treatment for glioblastoma.
- Published
- 2013
- Full Text
- View/download PDF
33. Functional characterization of an isoform-selective inhibitor of PI3K-p110β as a potential anticancer agent.
- Author
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Ni J, Liu Q, Xie S, Carlson C, Von T, Vogel K, Riddle S, Benes C, Eck M, Roberts T, Gray N, and Zhao J
- Subjects
- Animals, Antineoplastic Agents pharmacology, Cell Line, Cell Line, Tumor, Class Ia Phosphatidylinositol 3-Kinase metabolism, Enzyme Inhibitors pharmacology, Female, Humans, Indazoles pharmacology, Indazoles therapeutic use, Male, Mice, Mice, Nude, Neoplasm Transplantation, Neoplasms metabolism, Neoplasms pathology, Oncogene Protein v-akt metabolism, PTEN Phosphohydrolase deficiency, Protein Isoforms antagonists & inhibitors, Protein Isoforms metabolism, Sulfonamides pharmacology, Sulfonamides therapeutic use, Tumor Burden drug effects, Antineoplastic Agents therapeutic use, Enzyme Inhibitors therapeutic use, Neoplasms drug therapy, Phosphoinositide-3 Kinase Inhibitors
- Abstract
Unlabelled: Genetic approaches have shown that the p110β isoform of class Ia phosphatidylinositol-3-kinase (PI3K) is essential for the growth of PTEN-null tumors. Thus, it is desirable to develop p110β-specific inhibitors for cancer therapy. Using a panel of PI3K isoform-specific cellular assays, we screened a collection of compounds possessing activities against kinases in the PI3K superfamily and identified a potent and selective p110β inhibitor: KIN-193. We show that KIN-193 is efficacious specifically in blocking AKT signaling and tumor growth that are dependent on p110β activation or PTEN loss. Broad profiling across a panel of 422 human tumor cell lines shows that the PTEN mutation status of cancer cells strongly correlates with their response to KIN-193. Together, our data provide the first pharmacologic evidence that PTEN-deficient tumors are dependent on p110β in animals and suggest that KIN-193 can be pursued as a drug to treat tumors that are dependent on p110β while sparing other PI3K isoforms., Significance: We report the first functional characterization of a p110β-selective inhibitor, KIN-193, that is efficacious as an antitumor agent in mice. We show that this class of inhibitor holds great promise as a pharmacologic agent that could be used to address the potential therapeutic benefit of treating p110β-dependent PTEN-deficient human tumors., (© 2012 AACR)
- Published
- 2012
- Full Text
- View/download PDF
34. Kinome-wide selectivity profiling of ATP-competitive mammalian target of rapamycin (mTOR) inhibitors and characterization of their binding kinetics.
- Author
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Liu Q, Kirubakaran S, Hur W, Niepel M, Westover K, Thoreen CC, Wang J, Ni J, Patricelli MP, Vogel K, Riddle S, Waller DL, Traynor R, Sanda T, Zhao Z, Kang SA, Zhao J, Look AT, Sorger PK, Sabatini DM, and Gray NS
- Subjects
- Adenosine Triphosphate, Ataxia Telangiectasia Mutated Proteins, Cell Cycle Proteins antagonists & inhibitors, Cell Cycle Proteins metabolism, Cell Line, Tumor, DNA-Binding Proteins antagonists & inhibitors, DNA-Binding Proteins metabolism, Dose-Response Relationship, Drug, Enzyme Inhibitors pharmacology, Humans, Kinetics, Mechanistic Target of Rapamycin Complex 1, Multiprotein Complexes, Phosphatidylinositol 3-Kinases metabolism, Phosphoinositide-3 Kinase Inhibitors, Phosphorylation drug effects, Protein Serine-Threonine Kinases antagonists & inhibitors, Protein Serine-Threonine Kinases metabolism, Proteins antagonists & inhibitors, Proteins metabolism, Proteomics methods, Transcription Factors antagonists & inhibitors, Transcription Factors metabolism, Tumor Suppressor Proteins antagonists & inhibitors, Tumor Suppressor Proteins metabolism, p38 Mitogen-Activated Protein Kinases antagonists & inhibitors, p38 Mitogen-Activated Protein Kinases metabolism, Enzyme Inhibitors pharmacokinetics, TOR Serine-Threonine Kinases antagonists & inhibitors, TOR Serine-Threonine Kinases metabolism
- Abstract
An intensive recent effort to develop ATP-competitive mTOR inhibitors has resulted in several potent and selective molecules such as Torin1, PP242, KU63794, and WYE354. These inhibitors are being widely used as pharmacological probes of mTOR-dependent biology. To determine the potency and specificity of these agents, we have undertaken a systematic kinome-wide effort to profile their selectivity and potency using chemical proteomics and assays for enzymatic activity, protein binding, and disruption of cellular signaling. Enzymatic and cellular assays revealed that all four compounds are potent inhibitors of mTORC1 and mTORC2, with Torin1 exhibiting ∼20-fold greater potency for inhibition of Thr-389 phosphorylation on S6 kinases (EC(50) = 2 nM) relative to other inhibitors. In vitro biochemical profiling at 10 μM revealed binding of PP242 to numerous kinases, although WYE354 and KU63794 bound only to p38 kinases and PI3K isoforms and Torin1 to ataxia telangiectasia mutated, ATM and Rad3-related protein, and DNA-PK. Analysis of these protein targets in cellular assays did not reveal any off-target activities for Torin1, WYE354, and KU63794 at concentrations below 1 μM but did show that PP242 efficiently inhibited the RET receptor (EC(50), 42 nM) and JAK1/2/3 kinases (EC(50), 780 nM). In addition, Torin1 displayed unusually slow kinetics for inhibition of the mTORC1/2 complex, a property likely to contribute to the pharmacology of this inhibitor. Our results demonstrated that, with the exception of PP242, available ATP-competitive compounds are highly selective mTOR inhibitors when applied to cells at concentrations below 1 μM and that the compounds may represent a starting point for medicinal chemistry efforts aimed at developing inhibitors of other PI3K kinase-related kinases.
- Published
- 2012
- Full Text
- View/download PDF
35. Surveillance mechanism linking Bub1 loss to the p53 pathway.
- Author
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Gjoerup OV, Wu J, Chandler-Militello D, Williams GL, Zhao J, Schaffhausen B, Jat PS, and Roberts TM
- Subjects
- Aneuploidy, Antigens, Viral, Tumor metabolism, Cell Line, Cell Proliferation, Cellular Senescence, Cyclin-Dependent Kinase Inhibitor p21 metabolism, Fibroblasts cytology, Fibroblasts metabolism, Humans, Protein Binding, Protein Serine-Threonine Kinases, Retinoblastoma Protein metabolism, Simian virus 40 immunology, Spindle Apparatus metabolism, Time Factors, Protein Kinases deficiency, Tumor Suppressor Protein p53 metabolism
- Abstract
Bub1 is a kinase believed to function primarily in the mitotic spindle checkpoint. Mutation or aberrant Bub1 expression is associated with chromosomal instability, aneuploidy, and human cancer. We now find that targeting Bub1 by RNAi or simian virus 40 (SV40) large T antigen in normal human diploid fibroblasts results in premature senescence. Interestingly, cells undergoing replicative senescence were also low in Bub1 expression, although ectopic Bub1 expression in presenescent cells was insufficient to extend lifespan. Premature senescence caused by lower Bub1 levels depends on p53. Senescence induction was blocked by dominant negative p53 expression or depletion of p21(CIP1), a p53 target. Importantly, cells with lower Bub1 levels and inactivated p53 became highly aneuploid. Taken together, our data highlight a role for p53 in monitoring Bub1 function, which may be part of a more general spindle checkpoint surveillance mechanism. Our data support the hypothesis that Bub1 compromise triggers p53-dependent senescence, which limits the production of aneuploid and potentially cancerous cells.
- Published
- 2007
- Full Text
- View/download PDF
36. Simian virus 40 large T antigen targets the spindle assembly checkpoint protein Bub1.
- Author
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Cotsiki M, Lock RL, Cheng Y, Williams GL, Zhao J, Perera D, Freire R, Entwistle A, Golemis EA, Roberts TM, Jat PS, and Gjoerup OV
- Subjects
- Amino Acid Sequence, Animals, Mice, Molecular Sequence Data, Precipitin Tests, Protein Binding, Protein Serine-Threonine Kinases, Sequence Homology, Amino Acid, Two-Hybrid System Techniques, Antigens, Polyomavirus Transforming metabolism, Protein Kinases metabolism
- Abstract
The mitotic spindle checkpoint protein Bub1 has been found to be mutated at low frequency in certain human cancers characterized by aneuploidy. Simian virus 40 large T antigen efficiently immortalizes rodent cells and occasionally transforms them to tumorigenicity. T antigen can also cause genomic instability, inducing chromosomal aberrations and aneuploidy. Here, we report an interaction between Bub1 and T antigen. T antigen coimmunoprecipitates with endogenous Bub1 and Bub3, another component of the spindle checkpoint complex. Genetic analysis demonstrates that the interaction of T antigen with Bub1 is not required for immortalization but is closely correlated with transformation. T antigen induces an override of the spindle checkpoint dependent on Bub1 binding. This interaction with proteins of the spindle checkpoint machinery suggests another role for T antigen and provides insight into its ability to cause chromosomal aberrations, aneuploidy, and transformation.
- Published
- 2004
- Full Text
- View/download PDF
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