دورية أكاديمية
Massively parallel base editing to map variant effects in human hematopoiesis.
| العنوان: | Massively parallel base editing to map variant effects in human hematopoiesis. |
|---|---|
| المؤلفون: | Martin-Rufino JD; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; PhD Program in Biological and Biomedical Sciences, Harvard Medical School, Boston, MA 02115, USA., Castano N; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Pang M; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard-MIT Health Sciences and Technology, Harvard Medical School, Boston, MA 02115, USA., Grody EI; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Joubran S; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Chemical Biology PhD Program, Harvard Medical School, Boston, MA 02115, USA., Caulier A; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Wahlster L; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Li T; Department of Pharmacology and Yale Cancer Biology Institute, Yale University School of Medicine, New Haven, CT 06510, USA., Qiu X; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA., Riera-Escandell AM; Independent Researcher, now at Google, Zurich, Switzerland., Newby GA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA., Al'Khafaji A; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Chaudhary S; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Black S; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Weng C; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA., Munson G; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Liu DR; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA; Howard Hughes Medical Institute, Harvard University, Cambridge, MA 02138, USA., Wlodarski MW; Department of Hematology, St Jude Children's Research Hospital, Memphis, TN 38105, USA., Sims K; St. Jude Affiliate Clinic at Our Lady of the Lake Children's Health, Baton Rouge, LA 70809, USA., Oakley JH; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta and Emory University, Atlanta, GA 30322, USA., Fasano RM; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta and Emory University, Atlanta, GA 30322, USA., Xavier RJ; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Center for Computational and Integrative Biology, Department of Molecular Biology, and Center for the Study of Inflammatory Bowel Disease, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA., Lander ES; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA., Klein DE; Department of Pharmacology and Yale Cancer Biology Institute, Yale University School of Medicine, New Haven, CT 06510, USA., Sankaran VG; Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA. Electronic address: sankaran@broadinstitute.org. |
| المصدر: | Cell [Cell] 2023 May 25; Vol. 186 (11), pp. 2456-2474.e24. Date of Electronic Publication: 2023 May 02. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Cell Press Country of Publication: United States NLM ID: 0413066 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1097-4172 (Electronic) Linking ISSN: 00928674 NLM ISO Abbreviation: Cell Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Cambridge, Ma : Cell Press Original Publication: Cambridge, MIT Press. |
| مواضيع طبية MeSH: | Gene Editing* , Hematopoietic Stem Cells*/metabolism, Humans ; Cell Differentiation ; CRISPR-Cas Systems ; Genome ; Hematopoiesis ; Genetic Engineering ; Single-Cell Analysis |
| مستخلص: | Systematic evaluation of the impact of genetic variants is critical for the study and treatment of human physiology and disease. While specific mutations can be introduced by genome engineering, we still lack scalable approaches that are applicable to the important setting of primary cells, such as blood and immune cells. Here, we describe the development of massively parallel base-editing screens in human hematopoietic stem and progenitor cells. Such approaches enable functional screens for variant effects across any hematopoietic differentiation state. Moreover, they allow for rich phenotyping through single-cell RNA sequencing readouts and separately for characterization of editing outcomes through pooled single-cell genotyping. We efficiently design improved leukemia immunotherapy approaches, comprehensively identify non-coding variants modulating fetal hemoglobin expression, define mechanisms regulating hematopoietic differentiation, and probe the pathogenicity of uncharacterized disease-associated variants. These strategies will advance effective and high-throughput variant-to-function mapping in human hematopoiesis to identify the causes of diverse diseases. Competing Interests: Declaration of interests D.R.L. and G.A.N. have filed patent applications on gene-editing technologies through the Broad Institute of MIT and Harvard. D.R.L. is a consultant and equity owner of Beam Therapeutics, Pairwise Plants, Prime Medicine, Chroma Medicine and Nvelop Therapeutics, companies that use or deliver genome editing or genome engineering technologies. R.J.X. is the co-founder of Jnana Therapeutics and Celsius Therapeutics, the director of Moonlake Immunotherapeutics and a scientific advisory board member to Nestle, all unrelated to the present work. V.G.S. serves as an advisor to and/or has equity in Branch Biosciences, Ensoma, Novartis, Forma, and Cellarity, all unrelated to the present work. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| التعليقات: | Comment in: Cell Rep Methods. 2023 Jul 24;3(7):100541. doi: 10.1016/j.crmeth.2023.100541. (PMID: 37533644) |
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| معلومات مُعتمدة: | R01 CA265726 United States CA NCI NIH HHS; T32 HL007574 United States HL NHLBI NIH HHS; K99 HL163805 United States HL NHLBI NIH HHS; RM1 HG009490 United States HG NHGRI NIH HHS; R35 GM118062 United States GM NIGMS NIH HHS; U01 AI142756 United States AI NIAID NIH HHS; R01 HL146500 United States HL NHLBI NIH HHS; R01 DK103794 United States DK NIDDK NIH HHS; U54 DK106829 United States DK NIDDK NIH HHS |
| فهرسة مساهمة: | Keywords: base editing; differentiation; functional screens; genome engineering; hematopoiesis; hematopoietic stem cell; primary cells; single-cell genomics |
| تواريخ الأحداث: | Date Created: 20230503 Date Completed: 20230531 Latest Revision: 20240619 |
| رمز التحديث: | 20240619 |
| مُعرف محوري في PubMed: | PMC10225359 |
| DOI: | 10.1016/j.cell.2023.03.035 |
| PMID: | 37137305 |
| قاعدة البيانات: | MEDLINE |
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