Myeloid Conditioning with c-kit-Targeted CAR-T Cells Enables Donor Stem Cell Engraftment
Myeloid Conditioning with c-kit-Targeted CAR-T Cells Enables Donor Stem Cell Engraftment
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DOI：
10.1016/j.ymthe.2018.03.003
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发表时间：
2018-05-02
影响因子：
12.4
通讯作者：
Malech, Harry L.
中科院分区：
文献类型：
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作者：
Arai, Yasuyuki;Choi, Uimook;Malech, Harry L.
我们报告了一种使用小鼠中的c-kit靶向嵌合抗原受体T（c-kit CAR-T）细胞进行骨髓（BM）调节的新方法。以前的报告使用抗c-kit或抗CD 45抗体连接到毒素如皂草素已经是有希望的。我们使用c-kit CAR-T细胞开发了一种截然不同的方法。最初的研究表明，c-kit CAR-T细胞在体外杀死造血干细胞，但在体内扩增较差，CAR-T细胞向BM中的迁移较差。用低剂量环磷酰胺（125 mg/kg）预处理受体小鼠以及CAR-T细胞中的CXCR 4转导增强了向BM的运输和在BM中的扩增（< 1%-13.1%）。这导致BM c-kit（+）群体显著耗竭（9.0%-0.1%）。由于在Thy1.2受体小鼠中使用了同源Thy1.1 CAR-T细胞，因此在供体BM移植之前，可以使用抗Thy1.1抗体在体内耗尽CAR-T细胞。这实现了20%-40%的多谱系植入。我们应用这种调节，通过野生型BM移植，使慢性肉芽肿病小鼠的平均矫正率达到28%。我们的研究结果提供了一个概念证明，即c-kit CAR-T细胞可以在没有化疗/放疗的情况下实现有效的BM调节。我们的工作还表明，运输受体的共表达可以增强CAR-T细胞对指定组织的靶向。
We report a novel approach to bone marrow (BM) conditioning using c-kit-targeted chimeric antigen receptor T (c-kit CAR-T) cells in mice. Previous reports using anti-c-kit or anti-CD45 antibody linked to a toxin such as saporin have been promising. We developed a distinctly different approach using c-kit CAR-T cells. Initial studies demonstrated in vitro killing of hematopoietic stem cells by c-kit CAR-T cells but poor expansion in vivo and poor migration of CAR-T cells into BM. Pre-treatment of recipient mice with low-dose cyclophosphamide (125 mg/kg) together with CXCR4 transduction in the CAR-T cells enhanced trafficking to and expansion in BM (< 1%-13.1%). This resulted in significant depletion of the BM c-kit(+) population (9.0%-0.1%). Because congenic Thy1.1 CAR-T cells were used in the Thy1.2-recipient mice, anti-Thy1.1 antibody could be used to deplete CAR-T cells in vivo before donor BM transplant. This achieved 20%-40% multilineage engraftment. We applied this conditioning to achieve an average of 28% correction of chronic granulomatous disease mice by wild-type BM transplant. Our findings provide a proof of concept that c-kit CAR-T cells can achieve effective BM conditioning without chemo-/radiotherapy. Our work also demonstrates that co-expression of a trafficking receptor can enhance targeting of CAR-T cells to a designated tissue.