@article{e1badeceace04c33af37851787b34d71,
title = "Genetic Inactivation of CD33 in Hematopoietic Stem Cells to Enable CAR T Cell Immunotherapy for Acute Myeloid Leukemia",
abstract = "The absence of cancer-restricted surface markers is a major impediment to antigen-specific immunotherapy using chimeric antigen receptor (CAR) T cells. For example, targeting the canonical myeloid marker CD33 in acute myeloid leukemia (AML) results in toxicity from destruction of normal myeloid cells. We hypothesized that a leukemia-specific antigen could be created by deleting CD33 from normal hematopoietic stem and progenitor cells (HSPCs), thereby generating a hematopoietic system resistant to CD33-targeted therapy and enabling specific targeting of AML with CAR T cells. We generated CD33-deficient human HSPCs and demonstrated normal engraftment and differentiation in immunodeficient mice. Autologous CD33 KO HSPC transplantation in rhesus macaques demonstrated long-term multilineage engraftment of gene-edited cells with normal myeloid function. CD33-deficient cells were impervious to CD33-targeting CAR T cells, allowing for efficient elimination of leukemia without myelotoxicity. These studies illuminate a novel approach to antigen-specific immunotherapy by genetically engineering the host to avoid on-target, off-tumor toxicity. Reconstitution of the immune system with CD33-negative human hematopoietic stem cells enables anti-CD33 CAR-T cell killing of acute myeloid leukemia while sparing myeloid development and function.",
keywords = "CD33, CRISPR/Cas9 gene editing, acute myeloid leukemia, chimeric antigen receptor T cells, hematopoiesis, immunotherapy, non-human primate hematopoiesis",
author = "Kim, {Miriam Y.} and Yu, {Kyung Rok} and Kenderian, {Saad S.} and Marco Ruella and Shirley Chen and Shin, {Tae Hoon} and Aljanahi, {Aisha A.} and Daniel Schreeder and Michael Klichinsky and Olga Shestova and Kozlowski, {Miroslaw S.} and Cummins, {Katherine D.} and Xinhe Shan and Maksim Shestov and Adam Bagg and Morrissette, {Jennifer J.D.} and Palak Sekhri and Lazzarotto, {Cicera R.} and Calvo, {Katherine R.} and Kuhns, {Douglas B.} and Donahue, {Robert E.} and Behbehani, {Gregory K.} and Tsai, {Shengdar Q.} and Dunbar, {Cynthia E.} and Saar Gill",
note = "Funding Information: We thank Peter Klein, Robert Vonderheide, and Carl June for constructive comments; Yangbing Zhao, Jiangtao Ren, and Jian Huang for technical assistance; Sam Sadigh for photomicroscopy; Amy Ziober and Li Ping for immunocytochemistry; Vania Aikawa for cytogenetic analysis; Taehyong Kim, Zhiping (Paul) Wang, and Jonathan Schug for assistance with analysis of NGS; Mary Sell and Sam Mignogno for providing human apheresis specimens; NHLBI DNA Sequencing and Genomics and the NHLBI Flow Cytometry Cores; Naoya Uchida and John Tisdale for providing control bone marrow; Mark Raffeld, Liqiang Xi, and Chingiz Underbayev for ddPCR consultation and technical assistance; Keyvan Keyvanfar, Stephanie Sellers, Lemlem Alemu, and Jun Zhu for technical assistance; Ilker Tunc for off-target prediction; Aylin Bonifacino for RM CD34+ immunoselections; and Allen Krouse, Mark Metzger, Sandra Price, and the veterinary staff for care of the macaques. This work was supported by the Leukemia & Lymphoma Society (SCOR grant; S.G.), the National Institute of Health ( 1 K08 CA194256-01 ; S.G.), the Korean Visiting Scientist Training Award (K.-R.Y.), and the Intramural Research Program of the NHLBI . Funding Information: We thank Peter Klein, Robert Vonderheide, and Carl June for constructive comments; Yangbing Zhao, Jiangtao Ren, and Jian Huang for technical assistance; Sam Sadigh for photomicroscopy; Amy Ziober and Li Ping for immunocytochemistry; Vania Aikawa for cytogenetic analysis; Taehyong Kim, Zhiping (Paul) Wang, and Jonathan Schug for assistance with analysis of NGS; Mary Sell and Sam Mignogno for providing human apheresis specimens; NHLBI DNA Sequencing and Genomics and the NHLBI Flow Cytometry Cores; Naoya Uchida and John Tisdale for providing control bone marrow; Mark Raffeld, Liqiang Xi, and Chingiz Underbayev for ddPCR consultation and technical assistance; Keyvan Keyvanfar, Stephanie Sellers, Lemlem Alemu, and Jun Zhu for technical assistance; Ilker Tunc for off-target prediction; Aylin Bonifacino for RM CD34+ immunoselections; and Allen Krouse, Mark Metzger, Sandra Price, and the veterinary staff for care of the macaques. This work was supported by the Leukemia & Lymphoma Society (SCOR grant; S.G.), the National Institute of Health (1 K08 CA194256-01; S.G.), the Korean Visiting Scientist Training Award (K.-R.Y.), and the Intramural Research Program of the NHLBI. Publisher Copyright: {\textcopyright} 2018 Elsevier Inc.",
year = "2018",
month = may,
day = "31",
doi = "10.1016/j.cell.2018.05.013",
language = "English",
volume = "173",
pages = "1439--1453.e19",
journal = "Cell",
issn = "0092-8674",
number = "6",
}