Regulation of lytic granule exocytosis in Natural Killer (NK) Cells

自然杀伤 (NK) 细胞中裂解颗粒胞吐作用的调节

• 批准号: 9566638
• 负责人: JOHN COLIGAN
• 金额: $ 11.01万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9566638
• 关键词: Abnormal Cell; Actins; Address; Animal Model; Antibodies; Apoptosis; Biological Assay; CHS protein; CHS1 gene; Cell Line; Cell model; Cell physiology; Cells; Clinical Management; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Cytoplasmic Granules; Cytoskeleton; Defect; Diagnostic; Disease; Exocytosis; Generations; Genes; Granzyme; Griscelli Syndrome; Host Defense; Human; Immune; Impairment; Individual; Investigation; Knowledge; Lead; Longitudinal Studies; Lymphocyte; Lymphocyte Biology; Lymphocyte Subset; Lysosomes; Lytic; Methods; Microtubules; Modernization; Molecular; Morphology; Movement; Mutation; Natural Killer Cells; Pathogenesis; Pathway interactions; Patients; Phenotype; Play; Proteins; Regulation; Regulator Genes; Risk; Role; Sampling; Severity of illness; Signal Transduction; Symptoms; Techniques; Transplantation; chediak-higashi syndrome; cytotoxic; cytotoxicity; disease mechanisms study; disease-causing mutation; histiocyte; killings; loss of function; microbial; pathogen; perforin; tool; trafficking; tumor

Natural killer (NK) cells comprise a subset of lymphocytes involved in protection against microbial pathogens and tumors. Because of their cytotoxic capacity, NK cells are able to directly eliminate abnormal cells. Lytic granules (secretory lysosomes) of NK cells, containing perforin and granzymes, are indispensable for NK-cell cytotoxicity because their release results in the induction of target-cell apoptosis. Defects in lytic granule secretion are associated with serious diseases, such as hemophagocytic lymphohistiocytosis and Chediak-Higashi, Hermansky-Pudlak, or Griscelli syndromes. Knowledge about the proteins involved in regulation of lytic granule release is very limited, and the granule-specific protein machinery involved in NK-cell exocytosis is poorly understood. Chediak-Higashi syndrome (CHS) is a rare lysosomal storage disorder caused by mutations in the lysosomal trafficking regulator gene, LYST. LYST encodes a 429 kDa protein with several distinct domains implicated in various aspects of vesicular trafficking: ARM/HEAT, PH, BEACH and WD-40, but its exact function remains to be elucidated. NK cells in CHS patients have been shown to have abnormal morphology and function. Nevertheless, our understanding of NK cell defects in CHS is limited and the exact role of LYST in cytotoxic lymphocyte biology is unknown. Therefore, this project emphasis has been to understand the role of LYST in regulation of human NK cell cytotoxicity. To address this issue, we disrupted LYST expression in human NK cell lines, and the effects of LYST loss-of-function on lytic granule composition, movement, exocytosis and NK cell cytotoxic potential were investigated. We have successfully generated a human cellular model of CHS that can be used to study consequences of LYST deficiency at a cellular level.

天然杀伤（NK）细胞包括涉及针对微生物病原体和肿瘤的淋巴细胞的子集。由于其细胞毒性能力，NK细胞能够直接消除异常细胞。 NK细胞中含有穿孔蛋白和颗粒酶的NK细胞的裂解颗粒（分泌溶酶体）对于NK细胞细胞毒性是必不可少的，因为它们的释放导致靶细胞细胞凋亡的诱导。裂解颗粒分泌的缺陷与严重的疾病有关，例如淋巴淋巴虫组织细胞增多症和切迪亚克·希加西（Chediak-Higashi），赫尔曼斯基·帕德拉克（Hermansky-Pudlak）或griscelli综合征。关于裂解颗粒释放调节的蛋白质的知识非常有限，并且对NK细胞胞吐作用涉及的颗粒特异性蛋白质机制也很少了解。 Chediak-Higashi综合征（CHS）是由溶酶体贩运调节剂基因突变引起的罕见的溶酶体储存障碍，Lyst。 LYST编码一个429 kDa蛋白，其中有几个不同的域，其中涉及囊泡运输的各个方面：ARM/HEAT，PH，BEACH，BEACH和WD-40，但其确切的功能尚待阐明。 CHS患者的NK细胞已被证明具有异常的形态和功能。然而，我们对CHS中NK细胞缺陷的理解是有限的，LYST在细胞毒性淋巴细胞生物学中的确切作用尚不清楚。 因此，该项目的重点是了解LYST在调节人NK细胞细胞毒性中的作用。为了解决这个问题，我们研究了功能丧失对裂解颗粒组成，运动，胞吐作用和NK细胞细胞毒性潜力的影响。我们已经成功地产生了CHS的人类细胞模型，该模型可用于研究细胞水平的LYST缺乏症的后果。