Hematologic malignancies are driven by combinations of genetic lesions that have been difficult to model in human cells. We used CRISPR/Cas9 genome engineering of primary adult and umbilical cord blood CD34⁺ human hematopoietic stem and progenitor cells (HSPCs), the cells of origin for myeloid pre-malignant and malignant diseases, followed by transplantation into immunodeficient mice to generate genetic models of clonal hematopoiesis and neoplasia. Human hematopoietic cells bearing mutations in combinations of genes, including cohesin complex genes, observed in myeloid malignancies generated immunophenotypically defined neoplastic clones capable of long-term, multi-lineage reconstitution and serial transplantation. Employing these models to investigate therapeutic efficacy, we found that TET2 and cohesin-mutated hematopoietic cells were sensitive to azacitidine treatment. These findings demonstrate the potential for generating genetically defined models of human myeloid diseases, and they are suitable for examining the biological consequences of somatic mutations and the testing of therapeutic agents.

中文翻译：

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人类造血干细胞中基于CRISPR / Cas9的多重基因组编辑可模拟克隆性造血和骨髓瘤形成。

血液恶性肿瘤是由难以在人类细胞中建模的遗传损伤组合驱动的。我们使用了CRISPR / Cas9基因组工程改造了原代成人和脐带血CD34 ⁺人类造血干细胞和祖细胞（HSPC），是髓样恶性和恶性疾病的起源细胞，然后移植到免疫缺陷小鼠中以产生克隆性造血和瘤形成的遗传模型。在骨髓恶性肿瘤中观察到的人类造血细胞带有基因结合，包括黏附素复合基因的突变，产生了能够长期，多谱系重组和连续移植的免疫表型定义的肿瘤克隆。利用这些模型来研究治疗功效，我们发现TET2和黏附素突变的造血细胞对阿扎胞苷治疗敏感。这些发现证明了产生人类骨髓疾病遗传定义模型的潜力，