KIR and HLA in cis and trans cooperatively shape human NK education

顺式和反式的 KIR 和 HLA 合作塑造人类 NK 教育

• 批准号: 9160652
• 负责人: KATHARINE C HSU
• 金额: $ 42.85万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-03 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9160652
• 关键词: Acute leukemia; Affinity; Allogenic; Allografting; Binding; Bone Marrow; CD34 gene; Cell Therapy; Cell Transplants; Cell surface; Cells; Chimera organism; Development; Disease; Donor Selection; Education; Energy Transfer; Engraftment; Environment; Evaluation; Exhibits; Exposure to; Failure; Gene Family; Genetic; Goals; HLA Antigens; Hematologic Neoplasms; Hematopoietic; Hematopoietic Stem Cell Transplantation; Human; Immune; Immune system; Immunity; Immunogenetics; Immunology; In Vitro; Inflammatory; Investigation; Knowledge; Licensing; Ligands; Ligation; Lymphocyte; Mediating; Membrane; Mission; Molecular; Monoclonal Antibodies; Mouse Strains; Mus; Natural Killer Cells; Patients; Play; Population; Proteins; Public Health; RNA Interference; Relapse; Research; Role; Sampling; Shapes; Solid; Stem cells; Study Subject; System; Technology; Testing; Transgenic Mice; Transgenic Organisms; Transplant Recipients; Transplantation; Umbilical Cord Blood; Virus; Virus Diseases; cancer cell; cancer therapy; clinically relevant; cytotoxicity; hematopoietic cell transplantation; humanized mouse; in vivo; in vivo Model; killings; leukemia; mouse model; non-Native; novel; programs; receptor; relapse risk; response; therapy design; tumor

PROJECT SUMMARY Natural Killer (NK) cells are highly relevant in the setting of hematopoietic stem cell transplantation (HCT), where their activity can drastically reduce the risk of relapse in leukemia patients. Despite advances in donor selection to minimize transplant complications, leukemia relapse remains one of the most devastating causes of transplant failure. There is a general lack of knowledge of how NK cell populations are educated for effector function, leukemia recognition and clearance. While it is known that interaction between HLA class I molecules and the inhibitory killer Ig-like receptors (KIR) are important for NK education or “licensing” for effector function, the orientation of their interaction, the factors that contribute to the strength of their interaction, and the relative contributions of donor and recipient HLA are not known. A major impediment to fundamental human NK studies has been the lack of in vivo models that faithfully recapitulate NK education. A humanized NOD-RAG2IL2Rγc-/- mouse strain that exhibits HLA-B*27:05 successfully permits engraftment of mature NK cells and development of functional human NK cells from CD34+ stem cells. Preliminary investigations reveal that in vivo environments expressing cognate HLA educate developing and transferred KIR-expressing NK cells and enhance their long-term survival. In parallel, FRET and RNAi technology reveals that cis-interaction between HLA and KIR intrinsic to the cell also facilitates NK education. Evidence that HLA molecules can be transferred onto NK cells from adjacent cells provokes investigation of trogocytosis as a mechanism by which trans HLA may permit education via cis ligation of KIR. Restriction of HLA expression to the hematopoietic or stromal compartments in bone marrow chimeras will help define the relative contributions of donor and recipient HLA to NK licensing and persistence. Receptor- ligand pairs known to bind in trans also interact in cis, and further clarification is required to understand how these interactions influence NK function. Competition between cis-trans binding will test the hypothesis that cis interactions facilitate NK responsiveness by shielding KIR from ligation of trans HLA. NK cells with and without non-native HLA expression will be functionally tested to better understand how trogocytosis contributes to NK education after continuous or interrupted exposure to cognate HLA. Finally, experimental findings from the humanized mouse model and in vitro studies will be verified in a fully human system using NK cells obtained from patients who have received an HLA-mismatched umbilical cord blood transplant. These goals will advance our understanding of fundamental molecular requirements for NK education and acquisition of effector function. A deeper understanding of licensing by HLA provided in cis and trans will inform donor selection in hematopoietic cell transplantation and adoptive NK cell therapies for the treatment of cancer.

项目总结 自然杀伤(NK)细胞在造血干细胞移植(HCT)的设置中具有高度相关性， 他们的活动可以极大地降低白血病患者的复发风险。尽管捐赠者取得了进展 选择将移植并发症降至最低，白血病复发仍然是最具破坏性的原因之一 移植失败的可能性。人们普遍缺乏关于NK细胞群体是如何培养出效应器的知识。 功能、白血病识别和清除。虽然已知人类白细胞抗原I类分子之间的相互作用 抑制性杀伤分子Ig样受体(KIR)对NK教育或效应器功能的“许可”很重要， 他们相互作用的方向，影响他们相互作用强度的因素，以及相对的 供者和受者的人类白细胞抗原的贡献尚不清楚。 人类NK基础研究的一个主要障碍是缺乏体内模型 忠实地概括了NK教育。表现γ-B*27：05的人源化NOD-RAG2IL2RHLAC-/-小鼠株 成功地将成熟的NK细胞植入并从 CD34+干细胞。初步研究表明，在体内环境中表达同源人类白细胞抗原教育 开发和转移表达KIR的NK细胞，提高其长期存活率。同时，烦恼 RNAi技术表明，细胞固有的HLA和KIR之间的顺式相互作用也促进了NK 教育。有证据表明，人类白细胞抗原分子可以从相邻细胞转移到自然杀伤细胞上 巨噬细胞增多作为反式人类白细胞抗原允许通过顺式结扎KIR进行教育的机制的研究。 人类白细胞抗原在骨髓嵌合体中对造血室或基质室的限制表达 将有助于确定捐献者和接受者人类白细胞抗原对NK许可和持久性的相对贡献。受体- 已知的与反式结合的配体对也在顺式中相互作用，需要进一步澄清才能理解如何 这些相互作用影响NK功能。顺式-反式结合之间的竞争将检验顺式-反式结合的假设 相互作用通过保护KIR免受反式人类白细胞抗原的连接而促进NK的反应性。有无NK细胞 将对非原生人类白细胞抗原的表达进行功能测试，以更好地了解巨噬细胞增多是如何影响NK的 持续或间断暴露于同源人类白细胞抗原后的教育。最后，实验结果来自于 人源化小鼠模型和体外研究将在完全人类系统中使用获得的NK细胞进行验证 来自接受过人类白细胞抗原不相合脐带血移植的患者。 这些目标将促进我们对NK教育的基本分子要求的理解 以及效应器功能的获取。对顺式和转式遗嘱中的人类白细胞抗原授权有更深的理解 在造血细胞移植和过继NK细胞疗法治疗慢性粒细胞白血病中的供者选择 癌症。