Cancer cell:原来T细胞也会“衰老”,中国科学家揭示调节T细胞衰竭的关键激酶

2020-09-11 生物探索 生物探索

改善T细胞衰竭和增强效应子功能是增进免疫疗法的有效策略。

内源性或转移的细胞毒性T细胞是抗肿瘤免疫的基本介质,持续的抗原暴露会使T细胞逐渐变成衰竭状态。了解如何防止T细胞衰竭,从而扩展其功能是目前免疫肿瘤学中最紧迫的问题之一。?

造血祖细胞激酶1(HPK1)是一种免疫抑制调节激酶,也是一种T细胞受体(TCR)的负调节因子,会破坏TCR信号复合体的稳定性。先前的研究表明,HPK1激酶可以抑制多种细胞的免疫功能,而灭活其结构域足以引发抗肿瘤免疫反应效应。这表明,HPK1是一个很有前途的肿瘤免疫治疗的候选靶点。 ?

2020年8月28日,清华大学廖学斌课题组与中山大学魏来课题组合作, 揭示了HPK1介导T细胞功能障碍,并且是T细胞免疫疗法的药物靶标。相关成果发表在《Cancer cell》上。

在之前的研究中,研究人员已经证实,在25种不同类型癌症的肿瘤浸润性T细胞中,抑制性PDCD1(编码PD-1)受体和MAP4K1(编码HPK1)之间存在强烈的正相关关系。现通过进一步检测MAP4K1与肿瘤浸润性T细胞中的其他抑制性受体的相关性发现,不同癌症患者的MAP4K1与T细胞衰竭信号(CD3E、TIGIT、PDCD1、CTLA4、HAVCR2和LAG3)呈正相关,而检测患者肿瘤标本中抑制受体和HPK1的蛋白表达时也发现衰竭T细胞中HPK1表达上调。这些结果证实了HPK1与肿瘤浸润性T细胞耗竭呈正相关,提示HPK1可能是调节T细胞耗竭和抑制抗肿瘤免疫反应的关键激酶。?

不同癌症患者的MAP4K1表达及与T细胞衰竭信号相关性 

为了进一步验证HPK1对T细胞的抗肿瘤活性或肿瘤细胞生长的影响,研究人员检测了HPK1在肿瘤细胞中的表达,发现HPK1在肿瘤细胞中很少表达。研究人员采用B16OVA模型,发现与野生小鼠相比,MAP4K1细胞中OVA特异性CD8+肿瘤浸润淋巴细胞(TILs)的数量增加,MAP4K1缺陷小鼠表现出脱颗粒标记物CD107a的表达增加,并在佛波醇PMA和离子霉素刺激下产生更多的颗粒酶B和干扰素-g(IFN-g)。因此, MAP4K1缺失的CD8+TILs不仅能减少衰竭,而且具有较强的抗肿瘤活性。

之后研究人员进行相关机制研究,发现HPK1-Blimp1轴可驱动CD8+直到耗尽。研究人员先分析了肿瘤浸润性T细胞的RNA序列数据,发现HPK1基因敲除引起共抑制受体(如PDCD1、HAVCR2、LAG-3和CTLA-4)和凋亡相关基因(如ANXA1、FAS、BCL2L2、CASP3、CASP1和CASP4)的下调,细胞周期相关基因的上调(例如,GADD45B、CDKN2A、CDK2、CCNB1、MAD1L1、TFDP1、CDC25B和CDC25C),而小鼠体内蛋白表达水平也证实了该结果。 

此外,在黑色素瘤患者中,PRDM1(编码Blimp 1)和TOX与MAP4K1呈显着正相关。先前的研究证明,HPK1作为一种关键的TCR近端激酶发挥作用,其通过阳性细胞调节肿瘤浸润性T细胞衰竭Blimp1表达对T细胞活化进行调控,而本研究中的一系列实验也证实了这一结果。

HPK1-Blimp1轴及HPK1与其他信号通路之间的相互作用 

之后,研究人员进一步探讨了HPK1对嵌合抗原受体(CAR-T)细胞功能的影响。他们发现,与敲除了PD-1的 CAR-T细胞相比, 敲除MAP4K1的 CAR-T细胞对肿瘤细胞的毒性显着增强。 

接下来,研究人员采用患者衍生的MM NSG小鼠模型,评估MAP4K1敲除的CAR-T细胞的临床前疗效和安全性。结果显示,与对照组相比,在体外培养的第7天,MAP4K1敲除的B细胞成熟抗原CAR-T细胞中TOX和GATA3的表达降低,TCF1的表达增加。此外,其可显着抑制肿瘤生长并延长小鼠存活期。与对照组相比,这种细胞显着产生的并不是更高的IL-6,而是更高的IFN-g和更少的IL-15,而且毒性更小。这些结果表明,敲除MAP4K1的CAR-T细胞明显地平衡了安全性和有效性。

实验流程图 

鉴于?以上实验结果,研究人员开发了一系列HPK1的小分子抑制剂,最终发现ZYF0033对IC50小于10 nm的HPK1显示出良好的效价。?

在毒性研究中,ZYF0033也凸显出了较大的优势。在小鼠模型中, ZYF0033抑制肿瘤生长并导致肿瘤内DC、NK细胞和CD107A+CD8+T细胞浸润增加,但减少了调节性T细胞、PD-1+CD8+T细胞、TIM-3+CD8+T细胞和LAG3+CD8+T细胞的浸润。随后的药物化学优化开发出了蛋白水解靶向嵌合体(PROTAC)试剂SS47并进行了一系列的实验验证了其安全性和有效性。?

总结而言,遗传敲除、药理学抑制或PROTAC介导的HPK1降解在血液和实体瘤的各种临床前小鼠模型中提高了CAR-T细胞免疫疗法的功效。这些策略比在CAR-T细胞中遗传敲除PD-1更有效。?

改善T细胞衰竭和增强效应子功能是增进免疫疗法的有效策略。因此,开发HPK1抑制剂或通过PROTACs降解HPK1可能是肿瘤免疫治疗研究的新前沿。?

原始出处:

Jingwen Si, et al. Hematopoietic Progenitor Kinase1 (HPK1) Mediates T Cell Dysfunction and Is a Druggable Target for T Cell-Based Immunotherapies. Cancer Cell, Available online 28 August 2020.

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topicName=调节T细胞)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=aa5129877228, createdName=zhanfl, createdTime=Thu Jul 22 14:37:20 CST 2021, time=2021-07-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1962268, encodeId=811819622682b, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=395f25, createdName=爆笑小医, createdTime=Wed Apr 28 13:37:20 CST 2021, time=2021-04-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=2024384, encodeId=2fc520243846a, content=<a href='/topic/show?id=79f4219392b' target=_blank style='color:#2F92EE;'>#中国科学#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, 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ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885418, encodeId=595c885418b9, content=<a href='/topic/show?id=46ce580e865' target=_blank style='color:#2F92EE;'>#新冠肺炎#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=69, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=58078, encryptionId=46ce580e865, topicName=新冠肺炎)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Tue Sep 15 11:05:24 CST 2020, time=2020-09-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885110, encodeId=11b988511055, content=谢谢了, beContent=null, objectType=article, channel=null, level=null, likeNumber=74, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Mon Sep 14 10:07:14 CST 2020, time=2020-09-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1496095, encodeId=017c149609533, content=<a href='/topic/show?id=9c516685eea' target=_blank style='color:#2F92EE;'>#激酶#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66857, encryptionId=9c516685eea, topicName=激酶)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=73449239400, createdName=redcrab, createdTime=Sun Sep 13 12:37:20 CST 2020, time=2020-09-13, status=1, ipAttribution=)]
    2021-06-16 XFFF~

    可以

    0

  4. [GetPortalCommentsPageByObjectIdResponse(id=1780794, encodeId=10121e807947e, content=<a href='/topic/show?id=517a4016ac' target=_blank style='color:#2F92EE;'>#cancer cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=27, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4016, encryptionId=517a4016ac, topicName=cancer cell)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=96f62500201, createdName=12498e67m28暂无昵称, createdTime=Thu Dec 24 22:37:20 CST 2020, time=2020-12-24, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1871968, encodeId=5c9418e1968e7, content=<a href='/topic/show?id=fd764459a8' target=_blank style='color:#2F92EE;'>#Cell#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=35, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4459, encryptionId=fd764459a8, topicName=Cell)], attachment=null, authenticateStatus=null, createdAvatar=, 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topicName=调节T细胞)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=aa5129877228, createdName=zhanfl, createdTime=Thu Jul 22 14:37:20 CST 2021, time=2021-07-22, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1962268, encodeId=811819622682b, content=<a href='/topic/show?id=dbe6445416' target=_blank style='color:#2F92EE;'>#CEL#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=4454, encryptionId=dbe6445416, topicName=CEL)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=395f25, createdName=爆笑小医, createdTime=Wed Apr 28 13:37:20 CST 2021, time=2021-04-28, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=2024384, encodeId=2fc520243846a, content=<a href='/topic/show?id=79f4219392b' target=_blank style='color:#2F92EE;'>#中国科学#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, 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  5. 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ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885418, encodeId=595c885418b9, content=<a href='/topic/show?id=46ce580e865' target=_blank style='color:#2F92EE;'>#新冠肺炎#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=69, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=58078, encryptionId=46ce580e865, topicName=新冠肺炎)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Tue Sep 15 11:05:24 CST 2020, time=2020-09-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885110, encodeId=11b988511055, content=谢谢了, beContent=null, objectType=article, channel=null, level=null, likeNumber=74, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Mon Sep 14 10:07:14 CST 2020, time=2020-09-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1496095, encodeId=017c149609533, content=<a href='/topic/show?id=9c516685eea' target=_blank style='color:#2F92EE;'>#激酶#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66857, encryptionId=9c516685eea, topicName=激酶)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=73449239400, createdName=redcrab, createdTime=Sun Sep 13 12:37:20 CST 2020, time=2020-09-13, status=1, ipAttribution=)]
    2021-04-28 爆笑小医
  6. 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  7. 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  8. 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  9. 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ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885418, encodeId=595c885418b9, content=<a href='/topic/show?id=46ce580e865' target=_blank style='color:#2F92EE;'>#新冠肺炎#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=69, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=58078, encryptionId=46ce580e865, topicName=新冠肺炎)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Tue Sep 15 11:05:24 CST 2020, time=2020-09-15, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=885110, encodeId=11b988511055, content=谢谢了, beContent=null, objectType=article, channel=null, level=null, likeNumber=74, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=b0825417521, createdName=ms6000000960504356, createdTime=Mon Sep 14 10:07:14 CST 2020, time=2020-09-14, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1496095, encodeId=017c149609533, content=<a href='/topic/show?id=9c516685eea' target=_blank style='color:#2F92EE;'>#激酶#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=25, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=66857, encryptionId=9c516685eea, topicName=激酶)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=73449239400, createdName=redcrab, createdTime=Sun Sep 13 12:37:20 CST 2020, time=2020-09-13, status=1, ipAttribution=)]
    2020-09-14 ms6000000960504356

    谢谢了

    0

  10. 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    2020-09-13 redcrab

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