title: Engineered Murine HSCs Reconstitute Multi-lineage Hematopoiesis and Adaptive Immunity.
abstract: Hematopoietic stem cell (HSC) transplantation is curative for malignant and genetic blood disorders, but is limited by donor availability and immune-mismatch. Deriving HSCs from patient-matched embryonic/induced-pluripotent stem cells (ESCs/iPSCs) could address these limitations. Prior efforts in murine models exploited ectopic HoxB4 expression to drive self-renewal and enable multi-lineage reconstitution, yet fell short in delivering robust lymphoid engraftment. Here, by titrating exposure of HoxB4-ESC-HSC to Notch ligands, we report derivation of engineered HSCs that self-renew, repopulate multi-lineage hematopoiesis in primary and secondary engrafted mice, and endow adaptive immunity in immune-deficient recipients. Single-cell analysis shows that following engraftment in the bone marrow niche, these engineered HSCs further specify to a hybrid cell type, in which distinct gene regulatory networks of hematopoietic stem/progenitors and differentiated hematopoietic lineages are co-expressed. Our work demonstrates engineering of fully functional HSCs via modulation of genetic programs that govern self-renewal and lineage priming.
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Yi-Fen
authors.initials: YF
authors.lastname: Lu
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA; Department of Biomedical Engineering, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Cahan
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Samantha
authors.initials: S
authors.lastname: Ross
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Sahalie
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA.
authors.forename: Patricia M
authors.initials: PM
authors.lastname: Sousa
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Pediatrics, University of Washington, Seattle, WA 98105, USA.
authors.forename: Brandon K
authors.initials: BK
authors.lastname: Hadland
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA.
authors.forename: Wenqing
authors.initials: W
authors.lastname: Cai
authors.affiliation: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Serrao
authors.affiliation: Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
authors.forename: Alan N
authors.initials: AN
authors.lastname: Engelman
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Pediatrics, University of Washington, Seattle, WA 98105, USA.
authors.forename: Irwin D
authors.initials: ID
authors.lastname: Bernstein
authors.affiliation: Stem Cell Transplantation Program, Division of Pediatric Hematology/Oncology, Howard Hughes Medical Institute, Children's Hospital Boston, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Cambridge, MA 02138, USA. Electronic address: george.daley@childrens.harvard.edu.
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
mesh_terms: D056704:Adaptive Immunity / Q000235:genetics*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics / Q000276:immunology; D000066673:Cell Self Renewal / Q000235:genetics; D053263:Gene Regulatory Networks / Q000235:genetics; D006410:Hematopoiesis / Q000235:genetics / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics* / Q000276:immunology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D051880:Receptors, Notch / Q000235:genetics / Q000276:immunology; D059010:Single-Cell Analysis; D014157:Transcription Factors / Q000235:genetics* / Q000276:immunology
chemical_list: D018398:Homeodomain Proteins; C513262:Hoxb4 protein, mouse; D051880:Receptors, Notch; D014157:Transcription Factors
keywords: ESC-HSC; HSC engineering; HoxB4; Notch; adaptive immunity; adult globin; hematopoietic stem cell; lineage-priming