A CRISPR screen targeting PI3K effectors identifies RASA3 as a negative regulator of LFA-1-mediated adhesion in T cells.
A CRISPR screen targeting PI3K effectors identifies RASA3 as a negative regulator of LFA-1-mediated adhesion in T cells.
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DOI：
10.1126/scisignal.abl9169
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发表时间：
2022-07-19
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
--
作者：
整合素淋巴细胞功能相关抗原1（LFA-1）通过与细胞间粘附分子1（ICAM-1）和ICAM-2相互作用，帮助协调T细胞的迁移、粘附和活化。LFA-1在趋化因子受体和T细胞受体（TCR）通过由内向外信号传导的接合期间被激活，这是一个部分由磷酸肌醇3-激酶（PI 3 K）及其产物磷脂酰肌醇-3，4，5-三磷酸（PIP 3）介导的过程。为了评估PI 3 K在LFA-1活化中的潜在作用，我们设计了靶向已知和潜在的PIP 3结合蛋白的CRISPR/单向导RNA文库，并筛选了对原代小鼠T细胞结合ICAM-1的能力的影响。我们鉴定了多种调节LFA-1与ICAM-1结合的蛋白，包括Rap 1和Ras GTP酶激活蛋白RASA 3。我们发现RASA 3抑制T细胞中的LFA-1活化，其表达在T细胞活化后迅速降低，并且其活性被PI 3 K抑制。T细胞中RASA 3的缺失导致Rap 1活化增加，淋巴结进出缺陷，以及小鼠对T依赖性免疫的反应受损。我们的研究结果揭示了RASA 3在T细胞迁移，稳态和功能中的关键作用。
The integrin lymphocyte function–associated antigen 1 (LFA-1) helps to coordinate the migration, adhesion, and activation of T cells through interactions with intercellular adhesion molecule 1 (ICAM-1) and ICAM-2. LFA-1 is activated during the engagement of chemokine receptors and the T cell receptor (TCR) through inside-out signaling, a process that is partially mediated by phosphoinositide 3-kinase (PI3K) and its product phosphatidylinositol-3,4,5-trisphosphate (PIP3). To evaluate potential roles of PI3K in LFA-1 activation, we designed a library of CRISPR/single guide RNAs targeting known and potential PIP3-binding proteins and screened for effects on the ability of primary mouse T cells to bind to ICAM-1. We identified multiple proteins that regulated the binding of LFA-1 to ICAM-1, including the Rap1 and Ras GTPase-activating protein RASA3. We found that RASA3 suppressed LFA-1 activation in T cells, that its expression was rapidly reduced upon T cell activation, and that its activity was inhibited by PI3K. Loss of RASA3 in T cells led to increased Rap1 activation, defective lymph node entry and egress, and impaired responses to T-dependent immunization in mice. Our results reveal a critical role for RASA3 in T cell migration, homeostasis, and function.
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影响因子：
--
作者：
Huang B;Johansen KH;Schwartzberg PL
通讯作者：
Schwartzberg PL
DOI：
10.1074/jbc.m116.746867
发表时间：
2017-02-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Battram AM;Durrant TN;Agbani EO;Heesom KJ;Paul DS;Piatt R;Poole AW;Cullen PJ;Bergmeier W;Moore SF;Hers I
通讯作者：
Hers I
影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
5.4
作者：
Ardouin, L;Bracke, M;Tybulewicz, VLJ
通讯作者：
Tybulewicz, VLJ
DOI：
10.1073/pnas.0911573106
发表时间：
2009-12-01
影响因子：
11.1
作者：
Gigoux, Mathieu;Shang, Jijun;Suh, Woong-Kyung
通讯作者：
Suh, Woong-Kyung