Genetic Inactivation of CD33 in Hematopoietic Stem Cells to Enable CAR T Cell Immunotherapy for Acute Myeloid Leukemia

Miriam Y. Kim, Kyung Rok Yu, Saad S. Kenderian, Marco Ruella, Shirley Chen, Tae Hoon Shin, Aisha A. Aljanahi, Daniel Schreeder, Michael Klichinsky, Olga Shestova, Miroslaw S. Kozlowski, Katherine D. Cummins, Xinhe Shan, Maksim Shestov, Adam Bagg, Jennifer J.D. Morrissette, Palak Sekhri, Cicera R. Lazzarotto, Katherine R. Calvo, Douglas B. KuhnsRobert E. Donahue, Gregory K. Behbehani, Shengdar Q. Tsai, Cynthia E. Dunbar, Saar Gill

Research output: Contribution to journalArticlepeer-review

131 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)1439-1453.e19
Issue number6
StatePublished - May 31 2018


  • CD33
  • CRISPR/Cas9 gene editing
  • acute myeloid leukemia
  • chimeric antigen receptor T cells
  • hematopoiesis
  • immunotherapy
  • non-human primate hematopoiesis

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology


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