PO.IM01.07 · 免疫学
筛选富含成熟亚群的脐带血来源NK细胞可优化基于CAR-NK细胞的抗肿瘤治疗
Selecting cord blood-derived NK cells enriched in mature subsets optimizes CAR-NK cell-based anti-tumor therapy
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：CAR修饰的自然杀伤（CAR NK）细胞疗法提供了一种有前景的"现货型"细胞疗法，具有良好的临床活性和有利的安全性。然而，临床疗效在不同脐带血（CB）供体之间存在差异。深入理解驱动这种差异的细胞和分子因素对于改善CAR NK疗法的制造和性能至关重要。
方法：我们分析了来自我们首个在人体开展的CB来源CAR19/IL-15 NK细胞治疗B细胞恶性肿瘤临床研究（NCT03056339）的产品。我们此前已表明供体选择影响临床结局，一些供体与更好的反应及更长的无进展生存和总生存相关，而另一些则与较差的结局相关。为了明确这种效应的细胞学基础，我们回到那些特定供体，并对其NK细胞在操作前以及使用与试验相同方案进行体外扩增后进行了整合的单细胞RNA和抗体衍生标签（ADT）测序。然后我们将供体内在状态和特定细胞亚群与临床反应相关联。
结果：无监督聚类基于CD16和CD161的表达鉴定出两个不同的NK细胞亚群。一个双阴性（DN）亚群（CD56 bright CD16⁻ CD161⁻）及其扩增后代显示关键活化受体和衔接分子的表达降低，与扩增后有限的效应功能特征一致。相反，双阳性（DP）亚群（CD56 dim CD16⁺ CD161⁺）维持了稳健的适应性样细胞毒性特征，其特点是更强的膜近端活化信号和高效应基因表达，并在整个扩增过程中持续存在。值得注意的是，从富含DP亚群的CB供体生成的试验CAR-NK细胞产品与卓越的临床获益相关，而DN亚群的富集则与较差的结局相关。
结论：我们的单细胞多组学分析揭示了CB-NK细胞内此前未被充分认识的供体内在多样性，这可能解释了CAR NK细胞产品间的功能异质性。这些结果为供体选择策略以及为提高未来同种异体CAR NK细胞疗法一致性和效力的靶向基因改良提供了机制基础。
查看英文原文 English abstract
Background: CAR-modified natural killer (CAR NK) cell therapy offers a promising "off-the-shelf" cell therapy, with promising clinical activity with a favorable safety profile. Clinical efficacy, however, varies between umbilical cord blood (CB) donors. A deeper understanding of the cellular and molecular factors driving this variability is essential to improve the manufacturing and performance of CAR NK therapies.
Methods: We analyzed products from our first-in-human clinical study of CB-derived CAR19/IL-15 NK cells in B cell malignancies (NCT03056339). We had shown that donor selection affected clinical outcome, with some donors linked to better responses and longer progression free and overall survival, and others to poorer outcomes. To define the cellular basis for this effect, we returned to those specific donors and performed integrated single-cell RNA and antibody-derived tag (ADT) sequencing on their NK cells both before manipulation and after ex vivo expansion using the same protocol used for the trial. We then related donor intrinsic states and specific cellular subsets with clinical responses.
Results: Unsupervised clustering identified two distinct NK cell subsets based on their expression of CD16 and CD161. A double-negative (DN) subset (CD56 bright CD16 - CD161 - ) and its expanded progeny showed reduced expression of key activating receptors and adaptor molecules, consistent with limited effector function signatures after expansion. Conversely, the double-positive (DP) subset (CD56 dim CD16 + CD161 + ) maintained a robust adaptive-like cytotoxic profile, characterized by stronger membrane-proximal activation signaling and high effector-gene expression that persisted through expansion. Notably, trial CAR-NK cell products generated from CB donors enriched for the DP subset were associated with superior clinical benefits, whereas enrichment for the DN subset was associated with worse outcomes.
Conclusions: Our single-cell multiomic analysis reveals previously underappreciated donor-intrinsic diversity within CB-NK cells, which may explain the functional heterogeneity among CAR NK cell products. These results provide a mechanistic foundation for donor selection strategies and for targeted genetic improvements that can raise the consistency and potency of future allogeneic CAR NK cell therapies.
利益披露 Disclosure
Y. Li, None..
W. Jeffrey, None.