Representative Disease Models Research Articles

GPCR signaling contributes to immune characteristics of microenvironment and process of EBV-induced lymphomagenesis

Epstein-Barr virus (EBV) is the oncogenic driver of multiple cancers. However, the underlying mechanism of virus-cancer immunological interaction during disease pathogenesis remains largely elusive. Here we reported the first comprehensive proteogenomic characterization of natural killer/T-cell lymphoma (NKTCL), a representative disease model to study EBV-induced lymphomagenesis, incorporating genomic, transcriptomic, and in-depth proteomic data. Our multi-omics analysis of NKTCL revealed that EBV gene pattern correlated with immune-related oncogenic signaling. Single-cell transcriptome further delineated the tumor microenvironment as immune-inflamed, -deficient, and -desert phenotypes, in association with different setpoints of cancer-immunity cycle. EBV interacted with transcriptional factors to provoke GPCR interactome (GPCRome) reprogramming. Enhanced expression of chemokine receptor-1 (CCR1) on malignant and immunosuppressive cells modulated virus-cancer interaction on microenvironment. Therapeutic targeting CCR1 showed promising efficacy with EBV eradication, T-cell activation, and lymphoma cell killing in NKTCL organoid. Collectively, our study identified a previously unknown GPCR-mediated malignant progression and translated sensors of viral molecules into EBV-specific anti-cancer therapeutics.

The Effects of Adiponectin on the Behavior of B-Cell Leukemia Cells: Insights from an In Vitro Study.

Background: Non-Hodgkin's lymphoma (NHL), the most frequent hematological neoplasm worldwide, represents a heterogeneous group of malignancies. The etiology of NHL remains to be fully elucidated, but the role of adipose tissue (AT) in immune function via the secretion of adipokines was recently recognized. Among adipokines, adiponectin has garnered attention for its beneficial properties. This study aimed to explore the in vitro effects of AdipoRon, an adiponectin agonist, on JVM-2, a lymphoblast cell line used as a representative disease model. Methods: JVM-2 cells were treated with different concentrations of AdipoRon to evaluate its effects on viability (via an MTT test), cell cycle distribution (via an FACS analysis), invasiveness (via a Matrigel assay) and colony-forming ability; protein expression was assessed via a real-time PCR (qPCR) and/or Western blotting (WB). Results: We found that the prolonged exposure of JVM-2 cells to AdipoRon led to a reduction in their viability due to a cytostatic effect. Additionally, AdipoRon stimulated both the formation of cell colonies and the expression of E-cadherin. Interestingly, the administration of AdipoRon increased the invasive potential of JVM-2 cells. Conclusions: Our findings indicate that adiponectin is involved in the regulation of different cellular processes of JVM-2 cells, supporting its potential association with a pro-tumorigenic phenotype and indicating that it might contribute to the increased aggressiveness and metastatic potential of B lymphoma cells. However, additional studies are required to fully understand the molecular mechanisms of adiponectin's actions on lymphoblasts and whether it may represent a marker of disease.

A polycistronic transgene design for combinatorial genetic perturbations from a single transcript in Drosophila.

Experimental models that capture the genetic complexity of human disease and allow mechanistic explorations of the underlying cell, tissue, and organ interactions are crucial to furthering our understanding of disease biology. Such models require combinatorial manipulations of multiple genes, often in more than one tissue at once. The ability to perform complex genetic manipulations in vivo is a key strength of Drosophila, where many tools for sophisticated and orthogonal genetic perturbations exist. However, combining the large number of transgenes required to establish more representative disease models and conducting mechanistic studies in these already complex genetic backgrounds is challenging. Here we present a design that pushes the limits of Drosophila genetics by allowing targeted combinatorial ectopic expression and knockdown of multiple genes from a single inducible transgene. The polycistronic transcript encoded by this transgene includes a synthetic short hairpin cluster cloned within an intron placed at the 5' end of the transcript, followed by two protein-coding sequences separated by the T2A sequence that mediates ribosome skipping. This technology is particularly useful for modeling genetically complex diseases like cancer, which typically involve concurrent activation of multiple oncogenes and loss of multiple tumor suppressors. Furthermore, consolidating multiple genetic perturbations into a single transgene further streamlines the ability to perform combinatorial genetic manipulations and makes it readily adaptable to a broad palette of transgenic systems. This flexible design for combinatorial genetic perturbations will also be a valuable tool for functionally exploring multigenic gene signatures identified from omics studies of human disease and creating humanized Drosophila models to characterize disease-associated variants in human genes. It can also be adapted for studying biological processes underlying normal tissue homeostasis and development that require simultaneous manipulation of many genes.

SINGLE‐CELL RNA‐SEQ OF CLASSIC HAIRY CELL LEUKEMIA REVEALS DISEASE DRIVERS LINKED TO INTRINSIC TREATMENT RESISTANCE AND IDENTIFIES DUSP1 AS POTENTIAL NEW THERAPEUTIC TARGET

Introduction: Standard treatment with purine analogs such as cladribine (2-CdA) achieves excellent remissions in classic hairy cell leukemia (HCL), but up to 25% of patients relapse early and there is a non-diminishing risk of late relapse in complete responders. Even when targeting the recurrent genetic driver mutation BRAF V600E, HCL cells frequently persist in the bone marrow suggesting cooperating biologic alterations facilitating disease survival and persistence. Methods: Single-cell RNA sequencing (scRNA-Seq) was performed in primary HCL cells derived from three long-term (>10 years progression-free survival, PFS) and three short-term responders after 2-CdA treatment (SR, <3 years PFS) and from up to three time points (HCL diagnosis, first, and second relapse). Functional studies were performed using the BRAF D594E-mutated HAIR-M cell line after validating BRAF D594E as an activating mutation with similar downstream signaling and inhibitory potential with BRAF inhibitors as compared to BRAF V600E in primary HCL cells. Results: scRNA-Seq revealed distinct HCL cell clusters characterized by marked overexpression of DUSP1, FOS and JUND in all six patients that were outgrowing at first and second relapse in all three SR. In these DUSP1hi/FOShi/JUNDhi HCL cells, PROGENy cancer pathway analysis demonstrated induced activation of MAPK pathways and gene ontology analysis found negative regulation of the pro-apoptotic p38-MAPK-cascade as potential underlying biologic alteration. BRAF D594E-mutated HAIR-M cells were selected as substitute for primary HCL cells in cell culture due to comparable changes upon exposure to BRAF inhibitors in BRAF kinase activity conformation as assessed by the modular kinase conformation biosensor platform (KinCon) and similar reduction in ERK phosphorylation levels in HEK293T cells transiently overexpressing either flag-tagged BRAF D594E or BRAF V600E. In HAIR-M, we confirm stable expression levels of DUSP1, FOS and JUND, which were highly increased when co-cultured with HS5 stromal cells. Whereas increased expression of FOS and JUND was reversible upon exposure to BRAF inhibitors, expression of DUSP1, a direct target of BRAF-MEK-ERK signaling, and phosphorylation of p38 remained unaffected. As DUSP1 preferentially dephosphorylates/inactivates p38, we studied exposure of the DUSP1-inhibitor BCI in co-culture and found re-phosphorylated p38. Conclusion: DUSP1hi/FOShi/JUNDhi HCL cells may represent a distinct subset of HCL cells highly dependent on environmental cues with an inherently more resistant phenotype to treatment with 2-CdA or BRAF-inhibitors. Inhibition of DUSP1 may represent a novel therapeutic approach by effectively truncating HCL cells from extracellular pro-survival stimuli. Finally, our results suggest that BRAF D594E-mutated HAIR-M cells may serve as a representative disease model for functional studies. Keywords: bioinformatics, computational and systems biology, indolent non-Hodgkin lymphoma, tumor biology and heterogeneity No conflicts of interests pertinent to the abstract.

Abstract B008: Investigating the evolution of undifferentiated soft tissue sarcomas in a genetically engineered mouse model

Abstract Undifferentiated soft tissue sarcomas (USTS) are an aggressive class of sarcomas with few effective treatment options. Because of the rarity of these sarcomas and the paucity of representative disease models, little is known about how they develop. We used single cell transcriptomics to study the initiation and evolution of an USTS in an autochthonous genetically engineered mouse model. We profiled multiple stages of tumor development and identified intermediary cell states that define sarcomagenesis. Interestingly, we identify two discrete evolutionary pathways that arise in parallel in the mouse model. To determine the human relevance of our findings, we are developing machine learning approaches to compare cell states between the mouse model with single-cell transcriptomes that we generated from patient-derived xenografts. This cross-species comparison will uncover core USTS cell states and define their ancestral state transitions. Our ultimate goal is to identify essential molecular features of USTS, which may represent entry points for future therapeutic strategies. Citation Format: Jason E. Chan, Jonathan Rub, Carleigh Sussman, Olivera Grbovic-huezo, William D. Tap, Cristina Antonescu, Sam Singer, Doron Betel, Tuomas Tammela. Investigating the evolution of undifferentiated soft tissue sarcomas in a genetically engineered mouse model [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B008.

Central nervous system organoids for modeling neurodegenerative diseases.

Recent advances in induced pluripotent stem cell (iPSC) technology have allowed researchers to generate neurodegenerative disease-specific iPSCs and use the cells to derive a variety of relevant cell populations for laboratory modeling and drug testing. Nevertheless, these efforts have faced challenges related to immaturity and lack of complex developmental niches in the derived cell populations, limiting the utility of these in vitro models of neurodegenerative disease. Such limitations may be overcome by using human iPSC technology to generate three-dimensional (3D) brain organoids, which better recapitulate in vivo tissue architecture than traditional neuronal cultures to provide more complex and representative disease models and drug testing systems. In this review, we focus on the application of pluripotent stem cell-derived central nervous system (CNS) organoids to model neurodegenerative diseases. We first summarize recent progress in generating and characterizing various CNS organoids from pluripotent stem cells. We then review the application of CNS organoids for modeling several different human neurodegenerative diseases. We also describe several novel pathological mechanisms and drugs that were studied using patient iPSC-derived CNS organoids. Finally, we discuss remaining challenges and emerging opportunities for the use of 3D brain organoids for in vitro modeling of CNS development and neurodegeneration.

Abstract A09: Glypican-2 targeted CAR T cells designed to effectively eradicate endogenous site density solid tumors in the absence of toxicity

Abstract Background: Glypican-2 (GPC2) is a promising immunotherapeutic target, due to very low/absent expression on normal tissues (Bosse et al, Cancer Cell 2017). High cell surface GPC2 is found in numerous pediatric and adult malignancies, such as neuroblastoma, medulloblastoma, retinoblastoma, SCLC, and GBM. Here, we engineered GPC2-targeted CAR T-cells and optimized numerous modules (orientation of the variable heavy and light chain, hinge/transmembrane and co-stimulatory domains) to render CAR T cells highly efficacious against tumors expressing endogenous GPC2 site density in neuroblastoma xenograft models. Methods: GPC2 scFvs (GPC2.19 and GPC2.27) were isolated from a human Fab phage library and screened alongside with previously identified scFvs GPC2.D3 and GPC2.D4 in two orientations (N-terminal variable heavy or light chain) in second-generation retroviral vectors, possessing CD8a hinge/transmembrane (H/TM) and 41BB co-stimulatory domains. Potency was assessed in vitro for antigen-dependent cytokine production and killing and in vivo in neuroblastoma orthotopic subrenal capsule and/or flank PDX models. Results: While all constructs showed potent in vitro efficacy against isogenic target cells engineered to express GPC2 at supraphysiologic levels (GPC2Hi_Kelly-GPC2), efficacy against cell lines possessing endogenous site density (GPC2E_NBSD, SMS-SAN, KCNR) was modest. Based on cytokine production (IFNy, IL-2), killing capacity, and cross-reactivity against murine GPC2, GPC2.19VLVH was prioritized for in vivo studies, recapitulating in vitro findings. While CARs failed against GPC2E-models, significant antitumor effects were achieved against GPC2Hi, however, failing to cure. Target antigen density is an emerging determinant of CAR potency and our lab has recently demonstrated that incorporating CD28-H/TM and co-stimulatory domains exhibit advantages when targeting low site-density tumors (Majzner et al., ASH 2018). Strikingly, replacing H/TM domains derived from CD8a with those derived from CD28 in either 41BBζ or CD28ζ constructs resulted in a dramatic increase in potency and eradicated established GPC2E-tumors in orthotopic (NBSD) or PDX (COG-N-421x) neuroblastoma models, without signs of toxicity. While the majority of animals remained cured (&amp;gt;75 days), tumors relapsed in 30% of mice after a prolonged tumor-free interval. Principal component analysis (RNA-seq) of control-treated tumors and late relapses suggested strong clustering of transcriptional profiles based on these phenotypes and relapses were associated with ultralow protein and/or gene expression of GPC2 and other NB tumor antigens. Conclusion: We demonstrate that rational design of GPC2 CAR T-cells results in potent preclinical activity in representative disease models, laying the groundwork for clinical trials and establishing a model to shed light on tumor escape mechanisms after GPC2 CAR T-cell immune pressure. Citation Format: Sabine Heitzeneder, Kristopher R. Bosse, Zhongyu Zhu, Robbie G. Majzner, Johanna Theruvath, Peng Xu, Shaurya Dhingra, Hima Anbunathan, Anya Alag, Dimiter S. Dimitrov, John M. Maris, Crystal L. Mackall. Glypican-2 targeted CAR T cells designed to effectively eradicate endogenous site density solid tumors in the absence of toxicity [abstract]. In: Proceedings of the AACR Special Conference on the Advances in Pediatric Cancer Research; 2019 Sep 17-20; Montreal, QC, Canada. Philadelphia (PA): AACR; Cancer Res 2020;80(14 Suppl):Abstract nr A09.

SLC44A2 deficient mice have a reduced response in stenosis but not in hypercoagulability driven venous thrombosis

BackgroundGenome wide association studies (GWAS) identified SLC44A2 as a novel susceptibility gene for venous thrombosis (VT) and previous work established that SLC44A2 contributed to clot formation upon vascular injury. ObjectiveTo further investigate the role of SLC44A2 in VT by utilizing SLC44A2 deficient mice (Slc44a2−/−) in two representative disease models. MethodsMice were included in a hypercoagulability model driven by siRNA‐mediated hepatic gene silencing of anticoagulants Serpinc1 (antithrombin) and Proc (protein C) and a flow restriction (stenosis) model induced by partial ligation of the inferior vena cava. ResultsIn the hypercoagulability model, no effect in onset was observed in Slc44a2−/− animals; however, a drop in plasma fibrinogen and von Willebrand factor coinciding with an increase in blood neutrophils was recorded. In the neutrophil dependent stenosis model after 48 hours, Slc44a2−/− mice had significantly smaller thrombi both in length and weight with less platelet accumulation as a percentage of the total thrombus area. During the initiation of thrombosis at 6 hours post‐stenosis, Slc44a2−/− mice also had smaller thrombi both in length and weight, with circulating platelets remaining elevated in Slc44a2−/− animals. Platelet activation and aggregation under both static‐ and venous and arterial shear conditions were normal for blood from Slc44a2−/− mice. ConclusionsThese studies corroborate the original GWAS findings and establish a contributing role for SLC44A2 during the initiation of VT, with indications that this may be related to platelet‐neutrophil interaction. The precise mechanism however remains elusive and warrants further investigation.

A novel 3D-Printed preferential posterior mitral annular dilation device delineates regurgitation onset threshold in an ex vivo heart simulator

Mitral regurgitation (MR) due to annular dilation occurs in a variety of mitral valve diseases and is observed in many patients with heart failure due to mitral regurgitation. To understand the biomechanics of MR and ultimately design an optimized annuloplasty ring, a representative disease model with asymmetric dilation of the mitral annulus is needed. This work shows the design and implementation of a 3D-printed valve dilation device to preferentially dilate the posterior mitral valve annulus. Porcine mitral valves (n = 3) were sewn into the device and mounted within a left heart simulator that generates physiologic pressures and flows through the valves, while chordal forces were measured. The valves were incrementally dilated, inducing MR, while hemodynamic and force data were collected. Flow analysis demonstrated that MR increased linearly with respect to percent annular dilation when dilation was greater than a 25.6% dilation threshold (p < 0.01). Pre-threshold, dilation did not cause significant increases in regurgitant fraction. Forces on the chordae tendineae increased as dilation increased prior to the identified threshold (p < 0.01); post-threshold, the MR resulted in highly variable forces. Ultimately, this novel dilation device can be used to more accurately model a wide range of MR disease states and their corresponding repair techniques using ex vivo experimentation. In particular, this annular dilation device provides the means to investigate the design and optimization of novel annuloplasty rings.

Current Screening Methodologies in Drug Discovery for Selected Human Diseases.

The increase of many deadly diseases like infections by multidrug-resistant bacteria implies re-inventing the wheel on drug discovery. A better comprehension of the metabolisms and regulation of diseases, the increase in knowledge based on the study of disease-born microorganisms’ genomes, the development of more representative disease models and improvement of techniques, technologies, and computation applied to biology are advances that will foster drug discovery in upcoming years. In this paper, several aspects of current methodologies for drug discovery of antibacterial and antifungals, anti-tropical diseases, antibiofilm and antiquorum sensing, anticancer and neuroprotectors are considered. For drug discovery, two different complementary approaches can be applied: classical pharmacology, also known as phenotypic drug discovery, which is the historical basis of drug discovery, and reverse pharmacology, also designated target-based drug discovery. Screening methods based on phenotypic drug discovery have been used to discover new natural products mainly from terrestrial origin. Examples of the discovery of marine natural products are provided. A section on future trends provides a comprehensive overview on recent advances that will foster the pharmaceutical industry.

Validating traditional Chinese syndrome features in varied stages of chronic gastritis malignant transformation: study protocol for a cross-sectional study

IntroductionThe transition from chronic non-atrophic gastritis (CNAG) to chronic atrophic gastritis (CAG) and gastric carcinoma (GC) is regarded as a representative disease model of gastric mucosa malignant transformation led by...

Abstract LB-169: Asbestos and anthelmintics: Repurposing drugs to prevent mesothelioma in high-risk populations

Abstract The purpose of this study is to address the rising mortality rates in mesothelioma by using repurposed drugs in a chemoprevention setting. An initial screen of 100 repurposed agents at clinically relevant concentrations was conducted across three mesothelioma cell lines using AlamarBlue assays. Having identified the top three individual and combinatorial treatments, analysis of cell-cycle and apoptosis response to these drugs was used to corroborate the findings of the initial screen. As a more representative disease model a mouse PDX was developed from a human biphasic mesothelioma, and furthermore, primary tissue obtained, both of which we have used for explant experiments to assess the efficacy of the top treatments in a more realistic and translational model. As BAP-1 mutations are highly frequent in both acquired and spontaneous mesotheliomas, a CRISPR generated knock-out cell-line will model early-stage disease, and MET-5A mesothelial cells to assess normal cell response. The three top agents, comprising zinc acetate, niclosamide, and colchicine, have proven consistently efficacious in reducing mesothelioma cell-line viability by up to 72.59% individually and 93.55% in combination. Congruent results are provided by annexin V-FITC and FACS experiments which indicate treatments induce cell cycle alterations and apoptosis. Furthermore, FACS analysis of PDX explants also shows increased apoptosis of up to 27.0% in response to 24-hour treatment of two combined drugs. Importantly, the PDX still retains original histopathological features of the human tumour, making it an important foundation for further ex vivo and ultimately in vivo study. Initial results are promising and indicate that several repurposed agents significantly reduce survival and increase death of mesothelioma both in vitro and ex vivo. Mesothelioma, being predominantly caused by long-term asbestos exposure, has been under-researched due to an overreliance on the disuse of asbestos. As such, there is no effectual treatment and average survival remain static at under one year. However, following initial asbestos exposure, a 50-year latency precedes diagnosis, which offers an opportune window for a prevention strategy. Using the CRISPR-BAP-1-knockout cell-line to model early-stage disease, we hope to translate the top candidate into a primary prevention setting. However given the increasing frequency of late stage mesothelioma diagnoses, there is also a tangible need for secondary and tertiary prevention. Repurposed drugs, which offer long term safety and side effect profiles, as well as requiring less temporal and financial investment, allow expedited translation of any potential top candidate into clinic. Through further mechanistic analyses in in vitro and ex vivo cancer models, we hope to identify a single agent or combination that could significantly impede mesothelioma progression and ultimately reduce mortality. Citation Format: Bethan G. Rogoyski, Ankur Karmokar, Sameena Khan, Hong Cai, Farhat L. Khanim, Sara Busacca, Lynne Howells, Dean A. Fennell, Anne L. Thomas, Karen Brown. Asbestos and anthelmintics: Repurposing drugs to prevent mesothelioma in high-risk populations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-169.

Cardiopulmonary and histological characterization of an acute rat lung injury model demonstrating safety of mesenchymal stromal cell infusion

Background aimsIn the field of cellular therapy, potential cell entrapment in the lungs following intravenous administration in a compromised or injured pulmonary system is an important concern that requires further investigation. We developed a rat model of inflammatory and fibrotic lung disease to mimic the human clinical condition of obliterative bronchiolitis (OB) and evaluate the safety of intravenous infusion of mesenchymal stromal cells (MSCs). This model was used to obtain appropriate safety information and functional characterization to support the translation of an ex vivo–generated cellular product into human clinical trials. To overcome spontaneous recovery and size limitations associated with current animal models, we used a novel multiple dose bleomycin strategy to induce lasting lung injury in rats. MethodsIntratracheal instillation of bleomycin was administered to rats on multiple days. MSCs were intravenously infused 7 days apart. Detailed pulmonary function tests including forced expiratory volume, total lung capacity, and invasive hemodynamic measurements were conducted to define the representative disease model and monitor cardiopulmonary hemodynamic consequences of the cell infusion. Post-euthanasia assessments included a thorough evaluation of lung morphology and histopathology. ResultsThe double dose bleomycin instillation regimen resulted in severe and irreversible lung injury and fibrosis. Cardiopulmonary physiological monitoring reveled that no adverse events could be attributed to the cell infusion process. DiscussionAlthough our study did not show the infusion of MSCs to result in an improvement in lung function or rescue of damaged tissue this study does confirm the safety of MSC infusion into damaged lungs.

Mechanism of mitochondrial permeability transition pore induction and damage in the pancreas: inhibition prevents acute pancreatitis by protecting production of ATP

ObjectiveAcute pancreatitis is caused by toxins that induce acinar cell calcium overload, zymogen activation, cytokine release and cell death, yet is without specific drug therapy. Mitochondrial dysfunction has been implicated...

Towards understanding the underlying mechanism of vanishing white matter disease

s ingediend voor het Amsterdam Kindersymposium 2013 117 Towards understanding the underlying mechanism of vanishing white matter disease Lisanne E. Wisse, Emiel Polder, M. Clair van de Beek, Marjo S. van der Knaap, Truus E.M. Abbink Department of Pediatrics, Center for Childhood White Matter Disorders, VUmc INTRODUCTION Vanishing white matter disease (VWM) is one of the most prevalent genetic childhood white matter disorders. The disease is characterized by ataxia and spasticity. Diagnosis is performed by MRI, which shows progressive white matter degeneration, aggravated by stress, minor head trauma or fever. The lack of myelin and scar tissue formation in the aff ected area may result from a maturation defect in astrocytes and oligodendrocytes. VWM is caused by recessive mutations in the eukaryotic initiation factor 2B (eIF2B). This enzyme regulates the protein synthesis rate in every cell type and is regulated by various stressors, including heat shock. METHODS Our project aims to unravel the molecular mechanisms that underlie VWM. Although we have acquired increasing insights into the VWM brain pathology, it remains elusive how a defect in eIF2B causes a disease that almost exclusively aff ects the white matter. Aided by two clinically representative disease models, we currently address two unresolved questions. Firstly, we investigate whether relative white matter eIF2B levels deviate from those in other tissues using immunoblotting. Secondly, we address whether aberrant translation regulation of specifi c mRNAs in white matter cells underlies VWM using polysomal profi ling and pulsed isotope labeling. In follow-up assays we will assess the functional importance of the observed diff erences between control and VWM tissues. RESULTS Preliminary results suggest that we can purify polysomes from brain cells and tissue from normal and VWM mice. CONCLUSION Two representative disease models in combination with two robust genome-wide assays allow us to gain further insight into the disease mechanism of VWM.

A Blueprint for Advancing Genetics-Based Cancer Therapy

In the era of next-generation sequencing, there are significant challenges to harnessing cancer genome information to develop novel therapies. Key research thrusts in both academia and industry will speed this transition, and lessons learned for cancer will more broadly shape the process for genetic contributions to the therapy of disease more broadly.