Searched for: person:castep02
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The deubiquitinase USP9X regulates RIT1 protein abundance and oncogenic phenotypes
Riley, Amanda K; Grant, Michael; Snell, Aidan; Cromwell, Elizabeth; Vichas, Athea; Moorthi, Sitapriya; Rominger, Callie; Modukuri, Shrikar P; Urisman, Anatoly; Castel, Pau; Wan, Lixin; Berger, Alice H
PMID: 39161959
ISSN: 2589-0042
CID: 5680552
Functional and structural insights into RAS effector proteins
Mozzarelli, Alessandro M; Simanshu, Dhirendra K; Castel, Pau
RAS proteins are conserved guanosine triphosphate (GTP) hydrolases (GTPases) that act as molecular binary switches and play vital roles in numerous cellular processes. Upon GTP binding, RAS GTPases adopt an active conformation and interact with specific proteins termed RAS effectors that contain a conserved ubiquitin-like domain, thereby facilitating downstream signaling. Over 50 effector proteins have been identified in the human proteome, and many have been studied as potential mediators of RAS-dependent signaling pathways. Biochemical and structural analyses have provided mechanistic insights into these effectors, and studies using model organisms have complemented our understanding of their role in physiology and disease. Yet, many critical aspects regarding the dynamics and biological function of RAS-effector complexes remain to be elucidated. In this review, we discuss the mechanisms and functions of known RAS effector proteins, provide structural perspectives on RAS-effector interactions, evaluate their significance in RAS-mediated signaling, and explore their potential as therapeutic targets.
PMID: 39025071
ISSN: 1097-4164
CID: 5695942
Methylation of the chromatin modifier KMT2D by SMYD2 contributes to therapeutic response in hormone-dependent breast cancer
Blawski, Ryan; Vokshi, Bujamin H; Guo, Xinyu; Kittane, Srushti; Sallaku, Mirna; Chen, Wanlu; Gjyzari, Martina; Cheung, Tony; Zhang, Yuhan; Simpkins, Christopher; Zhou, Weiqiang; Kulick, Amanda; Zhao, Peihua; Wei, Meihan; Shivashankar, Pranavkrishna; Prioleau, Tatiana; Razavi, Pedram; Koche, Richard; Rebecca, Vito W; de Stanchina, Elisa; Castel, Pau; Chan, Ho Man; Scaltriti, Maurizio; Cocco, Emiliano; Ji, Hongkai; Luo, Minkui; Toska, Eneda
Activating mutations in PIK3CA are frequently found in estrogen-receptor-positive (ER+) breast cancer, and the combination of the phosphatidylinositol 3-kinase (PI3K) inhibitor alpelisib with anti-ER inhibitors is approved for therapy. We have previously demonstrated that the PI3K pathway regulates ER activity through phosphorylation of the chromatin modifier KMT2D. Here, we discovered a methylation site on KMT2D, at K1330 directly adjacent to S1331, catalyzed by the lysine methyltransferase SMYD2. SMYD2 loss attenuates alpelisib-induced KMT2D chromatin binding and alpelisib-mediated changes in gene expression, including ER-dependent transcription. Knockdown or pharmacological inhibition of SMYD2 sensitizes breast cancer cells, patient-derived organoids, and tumors to PI3K/AKT inhibition and endocrine therapy in part through KMT2D K1330 methylation. Together, our findings uncover a regulatory crosstalk between post-translational modifications that fine-tunes KMT2D function at the chromatin. This provides a rationale for the use of SMYD2 inhibitors in combination with PI3Kα/AKT inhibitors in the treatment of ER+/PIK3CA mutant breast cancer.
PMID: 38700982
ISSN: 2211-1247
CID: 5658222
The 8th International RASopathies Symposium: Expanding research and care practice through global collaboration and advocacy
Pierpont, Elizabeth I; Bennett, Anton M; Schoyer, Lisa; Stronach, Beth; Anschutz, April; Borrie, Sarah C; Briggs, Benjamin; Burkitt-Wright, Emma; Castel, Pau; Cirstea, Ion C; Draaisma, Fieke; Ellis, Michelle; Fear, Vanessa S; Frone, Megan N; Flex, Elisabetta; Gelb, Bruce D; Green, Tamar; Gripp, Karen W; Khoshkhoo, Sattar; Kieran, Mark W; Kleemann, Karolin; Klein-Tasman, Bonita P; Kontaridis, Maria I; Kruszka, Paul; Leoni, Chiara; Liu, Clifford Z; Merchant, Nadia; Magoulas, Pilar L; Moertel, Christopher; Prada, Carlos E; Rauen, Katherine A; Roelofs, Renée; Rossignol, Rodrigue; Sevilla, Christine; Sevilla, Gigi; Sheedy, Ryan; Stieglitz, Elliot; Sun, Daochun; Tiemens, Dagmar; White, Forest; Wingbermühle, Ellen; Wolf, Cordula; Zenker, Martin; Andelfinger, Gregor
Germline pathogenic variants in the RAS/mitogen-activated protein kinase (MAPK) signaling pathway are the molecular cause of RASopathies, a group of clinically overlapping genetic syndromes. RASopathies constitute a wide clinical spectrum characterized by distinct facial features, short stature, predisposition to cancer, and variable anomalies in nearly all the major body systems. With increasing global recognition of these conditions, the 8th International RASopathies Symposium spotlighted global perspectives on clinical care and research, including strategies for building international collaborations and developing diverse patient cohorts in anticipation of interventional trials. This biannual meeting, organized by RASopathies Network, was held in a hybrid virtual/in-person format. The agenda featured emerging discoveries and case findings as well as progress in preclinical and therapeutic pipelines. Stakeholders including basic scientists, clinician-scientists, practitioners, industry representatives, patients, and family advocates gathered to discuss cutting edge science, recognize current gaps in knowledge, and hear from people with RASopathies about the experience of daily living. Presentations by RASopathy self-advocates and early-stage investigators were featured throughout the program to encourage a sustainable, diverse, long-term research and advocacy partnership focused on improving health and bringing treatments to people with RASopathies.
PMID: 37969032
ISSN: 1552-4833
CID: 5610742
Papillomas of Costello syndrome are not associated with human papillomavirus infection in a small case series
Olsen, Gerilyn M; Johnson, Luke; Castel, Pau; Stevenson, David A; White, Kevin; Chiu, Yvonne E; Krol, Alfons; Siegel, Dawn H
PMID: 37028601
ISSN: 1097-6787
CID: 5502702
RAS-dependent RAF-MAPK hyperactivation by pathogenic RIT1 is a therapeutic target in Noonan syndrome-associated cardiac hypertrophy
Cuevas-Navarro, Antonio; Wagner, Morgan; Van, Richard; Swain, Monalisa; Mo, Stephanie; Columbus, John; Allison, Madeline R; Cheng, Alice; Messing, Simon; Turbyville, Thomas J; Simanshu, Dhirendra K; Sale, Matthew J; McCormick, Frank; Stephen, Andrew G; Castel, Pau
RIT1 is a RAS guanosine triphosphatase (GTPase) that regulates different aspects of signal transduction and is mutated in lung cancer, leukemia, and in the germline of individuals with Noonan syndrome. Pathogenic RIT1 proteins promote mitogen-activated protein kinase (MAPK) hyperactivation; however, this mechanism remains poorly understood. Here, we show that RAF kinases are direct effectors of membrane-bound mutant RIT1 necessary for MAPK activation. We identify critical residues in RIT1 that facilitate interaction with membrane lipids and show that these are necessary for association with RAF kinases and MAPK activation. Although mutant RIT1 binds to RAF kinases directly, it fails to activate MAPK signaling in the absence of classical RAS proteins. Consistent with aberrant RAF/MAPK activation as a driver of disease, we show that pathway inhibition alleviates cardiac hypertrophy in a mouse model of RIT1 mutant Noonan syndrome. These data shed light on the function of pathogenic RIT1 and identify avenues for therapeutic intervention.
PMCID:10348673
PMID: 37450595
ISSN: 2375-2548
CID: 5535322
Impaired proteolysis of non-canonical RAS proteins drives clonal hematopoietic transformation
Chen, Sisi; Vedula, Rahul S; Cuevas-Navarro, Antonio; Lu, Bin; Hogg, Simon J; Wang, Eric; Benbarche, Salima; Knorr, Katherine; Kim, Won Jun; Stanley, Robert F; Cho, Hana; Erickson, Caroline; Singer, Michael; Cui, Dan; Tittley, Steven; Durham, Benjamin H; Pavletich, Tatiana S; Fiala, Elise; Walsh, Michael F; Inoue, Daichi; Monette, Sebastien; Taylor, Justin; Rosen, Neal; McCormick, Frank; Lindsley, R Coleman; Castel, Pau; Abdel-Wahab, Omar
Recently, screens for mediators of resistance to FLT3 and ABL kinase inhibitors in leukemia resulted in the discovery of LZTR1 as an adaptor of a Cullin-3 RING E3 ubiquitin ligase complex responsible for degradation of RAS GTPases. In parallel, dysregulated LZTR1 expression via aberrant splicing and mutations were identified in clonal hematopoietic conditions. Here we identify that loss of LZTR1, or leukemia-associated mutants in the LZTR1 substrate and RAS GTPase RIT1 which escape degradation, drive hematopoietic stem cell (HSC) expansion and leukemia in vivo. While RIT1 stabilization was sufficient to drive hematopoietic transformation, transformation mediated by LZTR1 loss required MRAS. RAS targeting bioPROTACs or reduction of GTP-loaded RAS overcomes LZTR1 loss-mediated resistance to FLT3 inhibitors. These data reveal proteolysis of non-canonical RAS proteins as novel regulators of HSC self-renewal, define the function of RIT1 and LZTR1 mutations in leukemia, and identify means to overcome drug resistance due to LZTR1 downregulation.
PMID: 35904492
ISSN: 2159-8290
CID: 5276962
Noncanonical function of Capicua as a growth termination signal in Drosophila oogenesis
RodrÃguez-Muñoz, Laura; Lagares, Clà udia; González-Crespo, Sergio; Castel, Pau; Veraksa, Alexey; Jiménez, Gerardo
Capicua (Cic) proteins are conserved HMG-box transcriptional repressors that control receptor tyrosine kinase (RTK) signaling responses and are implicated in human neurological syndromes and cancer. While Cic is known to exist as short (Cic-S) and long (Cic-L) isoforms with identical HMG-box and associated core regions but distinct N termini, most previous studies have focused on Cic-S, leaving the function of Cic-L unexplored. Here we show that Cic-L acts in two capacities during Drosophila oogenesis: 1) as a canonical sensor of RTK signaling in somatic follicle cells, and 2) as a regulator of postmitotic growth in germline nurse cells. In these latter cells, Cic-L behaves as a temporal signal that terminates endoreplicative growth before they dump their contents into the oocyte. We show that Cic-L is necessary and sufficient for nurse cell endoreplication arrest and induces both stabilization of CycE and down-regulation of Myc. Surprisingly, this function depends mainly on the Cic-L-specific N-terminal module, which is capable of acting independently of the Cic HMG-box-containing core. Mirroring these observations, basal metazoans possess truncated Cic-like proteins composed only of Cic-L N-terminal sequences, suggesting that this module plays unique, ancient roles unrelated to the canonical function of Cic.
PMID: 35881788
ISSN: 1091-6490
CID: 5276362
The oncogenic PI3K-induced transcriptomic landscape reveals key functions in splicing and gene expression regulation
Ladewig, Erik; Michelini, Flavia; Jhaveri, Komal; Castel, Pau; Carmona, Javier; Fairchild, Lauren; Zuniga, Adler G; Arruabarrena-Aristorena, Amaia; Cocco, Emiliano; Blawski, Ryan; Kittane, Srushti; Zhang, Yuhan; Sallaku, Mirna; Baldino, Laura; Hristidis, Vasilis; Chandarlapaty, Sarat; Abdel-Wahab, Omar; Leslie, Christina; Scaltriti, Maurizio; Toska, Eneda
The PI3K pathway regulates proliferation, survival, and metabolism and is frequently activated across human cancers. A comprehensive elucidation of how this signaling pathway controls transcriptional and co-transcriptional processes could provide new insights into the key functions of PI3K signaling in cancer. Here, we undertook a transcriptomic approach to investigate genome-wide gene expression and transcription factor (TF) activity changes, as well as splicing and isoform usage dynamics, downstream of PI3K. These analyses uncovered widespread alternatively spliced (AS) isoforms linked to proliferation, metabolism, and splicing in PIK3CA mutant cells, which were reversed by inhibition of PI3Kα. Analysis of paired tumor biopsies from PIK3CA-mutated breast cancer patients undergoing treatment with PI3Kα inhibitors identified widespread splicing alterations that affect specific isoforms in common with the preclinical models, and these alterations, namely PTK2/FRNK and AFMID isoforms, were validated as functional drivers of cancer cell growth or migration. Mechanistically, isoform-specific splicing factors mediated PI3K-dependent RNA splicing. Treatment with splicing inhibitors rendered breast cancer cells more sensitive to the PI3Kα inhibitor alpelisib, resulting in greater growth inhibition than alpelisib alone. This study provides the first comprehensive analysis of widespread splicing alterations driven by oncogenic PI3K in breast cancer. The atlas of PI3K-mediated splicing programs establishes a key role for the PI3K pathway in regulating splicing, opening new avenues for exploiting PI3K signaling as a therapeutic vulnerability in breast cancer.
PMID: 35442400
ISSN: 1538-7445
CID: 5218342
The seventh international RASopathies symposium: Pathways to a cure-expanding knowledge, enhancing research, and therapeutic discovery
Kontaridis, Maria I; Roberts, Amy E; Schill, Lisa; Schoyer, Lisa; Stronach, Beth; Andelfinger, Gregor; Aoki, Yoko; Axelrad, Marni E; Bakker, Annette; Bennett, Anton M; Broniscer, Alberto; Castel, Pau; Chang, Caitlin A; Cyganek, Lukas; Das, Tirtha K; den Hertog, Jeroen; Galperin, Emilia; Garg, Shruti; Gelb, Bruce D; Gordon, Kristiana; Green, Tamar; Gripp, Karen W; Itkin, Maxim; Kiuru, Maija; Korf, Bruce R; Livingstone, Jeff R; López-Juárez, Alejandro; Magoulas, Pilar L; Mansour, Sahar; Milner, Theresa; Parker, Elisabeth; Pierpont, Elizabeth I; Plouffe, Kevin; Rauen, Katherine A; Shankar, Suma P; Smith, Shane B; Stevenson, David A; Tartaglia, Marco; Van, Richard; Wagner, Morgan E; Ware, Stephanie M; Zenker, Martin
RASopathies are a group of genetic disorders that are caused by genes that affect the canonical Ras/mitogen-activated protein kinase (MAPK) signaling pathway. Despite tremendous progress in understanding the molecular consequences of these genetic anomalies, little movement has been made in translating these findings to the clinic. This year, the seventh International RASopathies Symposium focused on expanding the research knowledge that we have gained over the years to enhance new discoveries in the field, ones that we hope can lead to effective therapeutic treatments. Indeed, for the first time, research efforts are finally being translated to the clinic, with compassionate use of Ras/MAPK pathway inhibitors for the treatment of RASopathies. This biannual meeting, organized by the RASopathies Network, brought together basic scientists, clinicians, clinician scientists, patients, advocates, and their families, as well as representatives from pharmaceutical companies and the National Institutes of Health. A history of RASopathy gene discovery, identification of new disease genes, and the latest research, both at the bench and in the clinic, were discussed.
PMID: 35266292
ISSN: 1552-4833
CID: 5182302