Individuals with age-related clonal hematopoiesis (CH) are at greater risk for hematologic malignancies and cardiovascular diseases. However, predictive preclinical animal models to recapitulate the spectrum of human CH are lacking. Through error-corrected sequencing of 56 human CH/myeloid malignancy genes, we identified natural CH driver mutations in aged rhesus macaques matching genes somatically mutated in human CH, with DNMT3A mutations being the most frequent. A CH model in young adult macaques was generated via autologous transplantation of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9-mediated gene-edited hematopoietic stem and progenitor cells (HSPCs), targeting the top human CH genes with loss-of-function (LOF) mutations. Long-term follow-up revealed reproducible and significant expansion of multiple HSPC clones with heterozygous TET2 LOF mutations, compared with minimal expansion of clones bearing other mutations. Although the blood counts of these CH macaques were normal, their bone marrows were hypercellular and myeloid-predominant. TET2-disrupted myeloid colony-forming units isolated from these animals showed a distinct hyperinflammatory gene expression profile compared with wild type. In addition, mature macrophages purified from the CH macaques showed elevated NLRP3 inflammasome activity and increased interleukin-1β (IL-1β) and IL-6 production. The model was used to test the impact of IL-6 blockage by tocilizumab, documenting a slowing of TET2-mutated expansion, suggesting that interruption of the IL-6 axis may remove the selective advantage of mutant HSPCs. These findings provide a model for examining the pathophysiology of CH and give insights into potential therapeutic interventions.

中文翻译：

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猕猴克隆造血模型展示了 TET2 破坏的克隆的扩展以及测试干预措施的效用。


患有与年龄相关的克隆造血（CH）的个体患血液系统恶性肿瘤和心血管疾病的风险更大。然而，缺乏概括人类慢性肝炎谱系的预测性临床前动物模型。通过对 56 个人类 CH/骨髓恶性肿瘤基因进行纠错测序，我们在老年恒河猴中发现了与人类 CH 体细胞突变基因相匹配的自然 CH 驱动突变，其中 DNMT3A 突变是最常见的。通过自体移植成簇规则间隔短回文重复序列 (CRISPR)/CRISPR 相关蛋白 9 介导的基因编辑造血干细胞和祖细胞 (HSPC)，在年轻成年猕猴中建立 CH 模型，靶向人类 CH 基因的丢失功能（LOF）突变。长期随访显示，与带有其他突变的克隆的最小扩增相比，具有杂合 TET2 LOF 突变的多个 HSPC 克隆具有可重复且显着的扩增。尽管这些 CH 猕猴的血细胞计数正常，但它们的骨髓细胞增多且以髓样细胞为主。与野生型相比，从这些动物中分离出的 TET2 破坏的骨髓集落形成单位显示出独特的高炎症基因表达谱。此外，从 CH 猕猴中纯化的成熟巨噬细胞显示出 NLRP3 炎性体活性升高，白细胞介素 1β (IL-1β) 和 IL-6 产量增加。该模型用于测试托珠单抗阻断 IL-6 的影响，记录了 TET2 突变扩增的减慢，表明 IL-6 轴的中断可能消除突变 HSPC 的选择性优势。这些发现为检查 CH 的病理生理学提供了一个模型，并为潜在的治疗干预提供了见解。