title: In Vivo Tracking of Human Hematopoiesis Reveals Patterns of Clonal Dynamics during Early and Steady-State Reconstitution Phases.
abstract: Hematopoietic stem/progenitor cells (HSPCs) are capable of supporting the lifelong production of blood cells exerting a wide spectrum of functions. Lentiviral vector HSPC gene therapy generates a human hematopoietic system stably marked at the clonal level by vector integration sites (ISs). Using IS analysis, we longitudinally tracked >89,000 clones from 15 distinct bone marrow and peripheral blood lineages purified up to 4 years after transplant in four Wiskott-Aldrich syndrome patients treated with HSPC gene therapy. We measured at the clonal level repopulating waves, populations' sizes and dynamics, activity of distinct HSPC subtypes, contribution of various progenitor classes during the early and late post-transplant phases, and hierarchical relationships among lineages. We discovered that in-vitro-manipulated HSPCs retain the ability to return to latency after transplant and can be physiologically reactivated, sustaining a stable hematopoietic output. This study constitutes in vivo comprehensive tracking in humans of hematopoietic clonal dynamics during the early and late post-transplant phases.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy. Electronic address: biasco.luca@hsr.it.
authors.forename: Luca
authors.initials: L
authors.lastname: Biasco
authors.affiliation: CUSSB, Vita-Salute University, 20132 Milan, Italy.
authors.forename: Danilo
authors.initials: D
authors.lastname: Pellin
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Scala
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Dionisio
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Basso-Ricci
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Lorena
authors.initials: L
authors.lastname: Leonardelli
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Samantha
authors.initials: S
authors.lastname: Scaramuzza
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Baricordi
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy; Pediatric Immunohematology and Bone Marrow Transplant Unit, San Raffaele Scientific Institute, 20132 Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Ferrua
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy; Pediatric Immunohematology and Bone Marrow Transplant Unit, San Raffaele Scientific Institute, 20132 Milan, Italy.
authors.forename: Maria Pia
authors.initials: MP
authors.lastname: Cicalese
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Giannelli
authors.affiliation: Target Sciences, GlaxoSmithKline R&D, Stevenage, Herts SG1 2NY, UK.
authors.forename: Victor
authors.initials: V
authors.lastname: Neduva
authors.affiliation: Target Sciences, GlaxoSmithKline R&D, Stevenage, Herts SG1 2NY, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Dow
authors.affiliation: National Center for Tumor Diseases (NCT), German Cancer Research Center (DKFZ), 69121 Heidelberg, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: National Center for Tumor Diseases (NCT), German Cancer Research Center (DKFZ), 69121 Heidelberg, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Von Kalle
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy; Division of Stem Cell Transplantation and Regenerative Medicine, Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Roncarolo
authors.affiliation: Pediatric Immunohematology and Bone Marrow Transplant Unit, San Raffaele Scientific Institute, 20132 Milan, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Ciceri
authors.affiliation: Department of Economy, University Roma Tre, 00154 Rome, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Vicard
authors.affiliation: Johann Bernoulli Institute, University of Groningen, 9700 AB Groningen, the Netherlands.
authors.forename: Ernst
authors.initials: E
authors.lastname: Wit
authors.affiliation: CUSSB, Vita-Salute University, 20132 Milan, Italy; Vita-Salute San Raffaele University, 20132 Milan, Italy.
authors.forename: Clelia
authors.initials: C
authors.lastname: Di Serio
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy; Vita-Salute San Raffaele University, 20132 Milan, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (TIGET), 20132 Milan, Italy; Pediatric Immunohematology and Bone Marrow Transplant Unit, San Raffaele Scientific Institute, 20132 Milan, Italy; Vita-Salute San Raffaele University, 20132 Milan, Italy. Electronic address: aiuti.alessandro@hsr.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Aiuti
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D060846:Cell Engineering; D019070:Cell Lineage / Q000235:genetics; D058948:Cell Tracking; D002675:Child, Preschool; D002999:Clone Cells; D015316:Genetic Therapy; D006410:Hematopoiesis / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007223:Infant; D008297:Male; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D016254:Mutagenesis, Insertional / Q000235:genetics; D013997:Time Factors; D014923:Wiskott-Aldrich Syndrome / Q000235:genetics / Q000628:therapy
chemical_list: D018952:Antigens, CD34