FATE1: Contributor to Tumor Cell Fitness and Example of an Oncogene-Activated Cancer Testis Antigen

Cancer Testis Antigens (CTAs) are a group of genes defined by their unique expression pattern, normally expressed nearly exclusively in gametogenic tissues they are also found aberrantly expressed in malignant tissues throughout the body. As a result of the immune-privileged nature of their normal site of expression in the testis, CTAs are capable of eliciting an immune response when expressed within tumors. As a result of their antigenicity, CTAs have been the intense studied as potential immunotherapy targets since their discovery in the early 1990s; however, in this time their possible functional contributions to tumorigenesis have been woefully under investigated. Here, we undertook the first comprehensive approach to define the functional contribution of CTAs to tumorigenesis. This screen identified numerous CTAs that support aspects of tumorigenesis. Further studies demonstrated that the uncharacterized CTA Fetal and Adult Testis-Expressed 1 (FATE1) is a major contributor to tumor cell fitness across multiple cancer lineages. We found that FATE1 is a mitochondrial protein that interacts with Mitochondrial Fission Factor (MFF) a mediator of mitochondrial fission and that expression of FATE1 is capable of altering mitochondrial morphology. Within the transformed cellular background, we find that FATE1 regulates protein levels of the tumor suppressor, Bcl-2 Interacting Killer (BIK), a pro-apoptotic member of the Bcl-2 protein family. We also found that BIK protein is degraded by the FATE1-interactor RNF183 a previously uncharacterized E3- ligase. We found further in vitro and clinical data that supports the hypothesis that FATE1 and RNF183 form a functionally relevant complex within tumors. Like their functional roles, the mechanisms by which CTAs are activated within tumors is currently unclear. Although demethylation plays a large role, epigenetic alterations alone are not sufficient to drive expression of all CTAs indicating that trans-acting factors are required. Here we find that FATE1 is a direct target of chimeric transcription factor EWS-FLI1, the oncogene responsible for the pediatric bone and soft tissue tumor Ewing sarcoma. FATE1 expression is required for short and long term viability of Ewing sarcoma derived cell lines. By leveraging a previously published EWS-FLI1 ChIP-seq dataset we nominate three additional CTAs (BORIS, MAGE-A4, and SPATA19) as EWS-FLI1 targets, two of which, MAGE-A4 and SPATA19 are required to maintain Ewing sarcoma cell viability. The results of these studies argue that not only do CTAs function to support tumor cell fitness but as targets of oncogenes they may potentially be key functional drivers in the early stages of tumorigenesis.Doctor of Philosoph

Mechanisms Promoting Escape from Mitotic Stress-Induced Tumor Cell Death

Non-small cell lung cancer (NSCLC) is notorious for its paltry responses to first-line therapeutic regimens. In contrast to acquired chemoresistance, little is known about the molecular underpinnings of the intrinsic resistance of chemo-naïve NSCLC. Here we report that intrinsic resistance to paclitaxel in NSCLC occurs at a cell-autonomous level due to the uncoupling of mitotic defects from apoptosis. To identify components that permit escape from mitotic stress-induced death, we employed a genome-wide RNAi-based strategy, which combines a high-throughput toxicity screen with a live-cell imaging platform to measure mitotic fate. This strategy revealed that prolonging mitotic arrest with a small molecule inhibitor of the APC/Cyclosome could sensitize otherwise paclitaxel-resistant NSCLC. We also defined novel roles for CASC1 and TRIM69 in supporting resistance to spindle poisons. CASC1, which is frequently co-amplified with KRAS in lung tumors, is essential for microtubule polymerization and satisfaction of the spindle assembly checkpoint. TRIM69, which associates with spindle poles and promotes centrosomal clustering, is essential for formation of a bipolar spindle. Notably, RNAi-mediated attenuation of CASC1 or TRIM69 was sufficient to inhibit tumor growth in vivo. On the basis of our results, we hypothesize that tumor evolution selects for a permissive mitotic checkpoint, which may promote survival despite chromosome segregation errors. Attacking this adaptation may restore the apoptotic consequences of mitotic damage to permit the therapeutic eradication of drug-resistant cancer cells

Comprehensive functional characterization of cancer–testis antigens defines obligate participation in multiple hallmarks of cancer

Tumours frequently activate genes whose expression is otherwise biased to the testis, collectively known as cancer–testis antigens (CTAs). The extent to which CTA expression represents epiphenomena or confers tumorigenic traits is unknown. In this study, to address this, we implemented a multidimensional functional genomics approach that incorporates 7 different phenotypic assays in 11 distinct disease settings. We identify 26 CTAs that are essential for tumor cell viability and/or are pathological drivers of HIF, WNT or TGFβ signalling. In particular, we discover that Foetal and Adult Testis Expressed 1 (FATE1) is a key survival factor in multiple oncogenic backgrounds. FATE1 prevents the accumulation of the stress-sensing BH3-only protein, BCL-2-Interacting Killer (BIK), thereby permitting viability in the presence of toxic stimuli. Furthermore, ZNF165 promotes TGFβ signalling by directly suppressing the expression of negative feedback regulatory pathways. This action is essential for the survival of triple negative breast cancer cells in vitro and in vivo. Thus, CTAs make significant direct contributions to tumour biology

A lab-on-a-chip system with an embedded porous membrane-based impedance biosensor array for nanoparticle risk assessment on placental Bewo trophoblast cells

The human placenta is a unique organ serving as the lung, gut, liver, and kidney of the fetus, mediating the exchange of different endogenous as well as exogenous substances and gases between the mother and fetus during pregnancy. Additionally, the placental barrier protects the fetus from a range of environmental toxins, bacterial and viral infections, since any contaminant bridging the placenta may have unforeseeable effects on embryonal and fetal development. A more recent concern in placenta research, however, involves the ability of engineered nanoparticles to cross the placental barrier and/or affect its barrier function. To advance nanoparticle risk assessment at the human placental barrier, we have developed as proof-of-principle a highly integrated placenta-on-a-chip system containing embedded membrane-bound impedance microsensor arrays capable of non-invasively monitoring placental barrier integrity. Barrier integrity is continuously and label-free evaluated using porous membrane-based interdigitated electrode structures located on top of a porous PET membrane supporting a barrier of trophoblast-derived BeWo cell barrier in the absence and presence of standardized silicon dioxide (SiO2), titanium dioxide (TiO2), and zinc oxide (ZnO) nanomaterials.This work has been funded by the European Union’s Horizon 2020 research and innovation program under grant agreement No. 685817.Peer reviewe

PAK6 phosphorylates 14-3-3γ to regulate steady state phosphorylation of LRRK2

Mutations in Leucine-rich repeat kinase 2 (LRRK2) are associated with Parkinson's disease (PD) and, as such, LRRK2 is considered a promising therapeutic target for age-related neurodegeneration. Although the cellular functions of LRRK2 in health and disease are incompletely understood, robust evidence indicates that PD-associated mutations alter LRRK2 kinase and GTPase activities with consequent deregulation of the downstream signaling pathways. We have previously demonstrated that one LRRK2 binding partner is P21 (RAC1) Activated Kinase 6 (PAK6). Here, we interrogate the PAK6 interactome and find that PAK6 binds a subset of 14-3-3 proteins in a kinase dependent manner. Furthermore, PAK6 efficiently phosphorylates 14-3-3γ at Ser59 and this phosphorylation serves as a switch to dissociate the chaperone from client proteins including LRRK2, a well-established 14-3-3 binding partner. We found that 14-3-3γ phosphorylated by PAK6 is no longer competent to bind LRRK2 at phospho-Ser935, causing LRRK2 dephosphorylation. To address whether these interactions are relevant in a neuronal context, we demonstrate that a constitutively active form of PAK6 rescues the G2019S LRRK2-associated neurite shortening through phosphorylation of 14-3-3γ. Our results identify PAK6 as the kinase for 14-3-3γ and reveal a novel regulatory mechanism of 14-3-3/LRRK2 complex in the brain

Search for new particles in events with energetic jets and large missing transverse momentum in proton-proton collisions at root s=13 TeV

A search is presented for new particles produced at the LHC in proton-proton collisions at root s = 13 TeV, using events with energetic jets and large missing transverse momentum. The analysis is based on a data sample corresponding to an integrated luminosity of 101 fb(-1), collected in 2017-2018 with the CMS detector. Machine learning techniques are used to define separate categories for events with narrow jets from initial-state radiation and events with large-radius jets consistent with a hadronic decay of a W or Z boson. A statistical combination is made with an earlier search based on a data sample of 36 fb(-1), collected in 2016. No significant excess of events is observed with respect to the standard model background expectation determined from control samples in data. The results are interpreted in terms of limits on the branching fraction of an invisible decay of the Higgs boson, as well as constraints on simplified models of dark matter, on first-generation scalar leptoquarks decaying to quarks and neutrinos, and on models with large extra dimensions. Several of the new limits, specifically for spin-1 dark matter mediators, pseudoscalar mediators, colored mediators, and leptoquarks, are the most restrictive to date.Peer reviewe

Measurement of the top quark forward-backward production asymmetry and the anomalous chromoelectric and chromomagnetic moments in pp collisions at √s = 13 TeV

Abstract The parton-level top quark (t) forward-backward asymmetry and the anomalous chromoelectric (d̂ t) and chromomagnetic (μ̂ t) moments have been measured using LHC pp collisions at a center-of-mass energy of 13 TeV, collected in the CMS detector in a data sample corresponding to an integrated luminosity of 35.9 fb−1. The linearized variable AFB(1) is used to approximate the asymmetry. Candidate t t ¯ events decaying to a muon or electron and jets in final states with low and high Lorentz boosts are selected and reconstructed using a fit of the kinematic distributions of the decay products to those expected for t t ¯ final states. The values found for the parameters are AFB(1)=0.048−0.087+0.095(stat)−0.029+0.020(syst),μ̂t=−0.024−0.009+0.013(stat)−0.011+0.016(syst), and a limit is placed on the magnitude of | d̂ t| < 0.03 at 95% confidence level. [Figure not available: see fulltext.

Combined searches for the production of supersymmetric top quark partners in proton-proton collisions at root s=13 TeV

A combination of searches for top squark pair production using proton-proton collision data at a center-of-mass energy of 13 TeV at the CERN LHC, corresponding to an integrated luminosity of 137 fb(-1) collected by the CMS experiment, is presented. Signatures with at least 2 jets and large missing transverse momentum are categorized into events with 0, 1, or 2 leptons. New results for regions of parameter space where the kinematical properties of top squark pair production and top quark pair production are very similar are presented. Depending on themodel, the combined result excludes a top squarkmass up to 1325 GeV for amassless neutralino, and a neutralinomass up to 700 GeV for a top squarkmass of 1150 GeV. Top squarks with masses from 145 to 295 GeV, for neutralino masses from 0 to 100 GeV, with a mass difference between the top squark and the neutralino in a window of 30 GeV around the mass of the top quark, are excluded for the first time with CMS data. The results of theses searches are also interpreted in an alternative signal model of dark matter production via a spin-0 mediator in association with a top quark pair. Upper limits are set on the cross section for mediator particle masses of up to 420 GeV

Probing effective field theory operators in the associated production of top quarks with a Z boson in multilepton final states at root s=13 TeV

Peer reviewe