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张锋教授Nature发文,CRISPR技术找到了癌症免疫疗法关键基因!

2017-08-08 佚名 药明康德

要说近年来癌症领域取得的最大突破,那莫过于免疫疗法了。一些原本等同于死刑判决的恶性癌症,在免疫疗法的作用下,竟能连续10多年不复发,这近乎达到了治愈的效果,堪称奇迹。 但免疫疗法也不是万能的。肿瘤细胞一直在快速突变,而有些突变会让免疫疗法失效。目前我们知道,B2M、JAK1与JAK2基因一旦发生突变失活,就会让免疫疗法失去应有的作用。

要说近年来癌症领域取得的最大突破,那莫过于免疫疗法了。一些原本等同于死刑判决的恶性癌症,在免疫疗法的作用下,竟能连续10多年不复发,这近乎达到了治愈的效果,堪称奇迹。

但免疫疗法也不是万能的。肿瘤细胞一直在快速突变,而有些突变会让免疫疗法失效。目前我们知道,B2M、JAK1与JAK2基因一旦发生突变失活,就会让免疫疗法失去应有的作用。

有趣的是,大自然有其“相生相克”的原理。一方面,肿瘤的突变可能会让免疫疗法失效;另一方面,肿瘤的突变,却能产生新的抗原,诱发免疫系统的攻击,提高免疫疗法的效果。因此,了解哪些基因突变可能会影响免疫疗法的效果(无论是提高还是抑制),对免疫疗法的开发非常重要。


▲我们找到了癌症免疫疗法的关键基因(图片来源:《自然》)

先前,CRISPR-Cas9基因组编辑技术已被用于筛选潜在的基因靶点。在癌症细胞中,通过筛选,我们能找到影响癌细胞增殖、产生耐药性、诱使转移的基因。一些科学家们就想到,我们应该可以用同样的技术,筛选出会影响免疫疗法的基因。

今天在线发表的一篇《自然》论文中,一群科学家们就找到了一个会影响免疫疗法的关键基因。本研究由美国国立卫生研究院(NIH)的Nicholas Restifo教授主导,著名华人学者,CRISPR技术先驱张锋教授协助参与。


▲本研究的通讯作者Nicholas Restifo教授(图片来源:《自然》)


▲张锋教授协助了这项研究(图片来源:MIT)

在这项研究中,科学家们开发了一种全新的“双细胞CRISPR”方法。该体系由人类的T细胞与黑色素瘤细胞组成。在癌症免疫疗法中,T细胞需要直接对癌细胞进行杀伤,因此两者间的结合非常重要。按科学家们的设想,如果某个基因突变会影响这两类细胞的结合,就说明它对免疫疗法很重要。

于是,科学家们建立了一个大型的筛选库,里面带有超过12万个sgRNA。这些sgRNA能靶向细胞内19050个编码蛋白质的基因,以及1864条微RNA。平均下来,每个基因上会被分配到6条sgRNA,每条微RNA上会被分配到4条sgRNA。这些sgRNA会帮助CRISPR-Cas9系统对这些基因或微RNA进行敲除,揭露出它们在免疫疗法中的作用。


▲“双细胞CRISPR”的设计(图片来源:《自然》)

科学家们发现,他们筛选出的基因,大部分与抗原递呈以及干扰素-γ的信号通路有关,这表明这些基因有望成为未来研究的重点。其中,科学家们对一个叫做APLNR的基因产生了兴趣。这个基因的多个突变都会影响到免疫疗法的效果,这一点也在不同的癌症细胞系中得到了证实。因此,它参与的信号通路,对于癌症免疫疗法的开发,有着直接的应用价值。

那么,APLNR基因究竟有啥作用呢?功能分析表明,它编码了一个G蛋白偶联受体。在许多癌症中,人们都观察到了它的突变。利用RNA测序的手段,科学家们在敲除了APLNR基因的细胞中分析了转录组,但与抗原递呈有关的mRNA转录水平没有发生明显变化。这表明APLNR可能是通过蛋白质的直接作用,影响了免疫疗法的效果。

在先前的数据库里,科学家们找到了线索。之前的一些数据表明APLNR可能会与JAK1蛋白结合,而我们知道JAK1蛋白会影响到免疫疗法。通过免疫沉淀实验,这一假设得到了验证。基于这些结果,科学家们提出了一个新机制:APLNR蛋白能结合JAK1,从而调节肿瘤对干扰素-γ的响应。如果它失去功能,就会降低免疫疗法的效果。与之相对应,如果过量表达APLNR,就能显着增强肿瘤细胞对T细胞攻击的敏感度。肿瘤细胞的细胞溶解率也从71.7%上升到了81.1%。换句话说,肿瘤免疫疗法的效果有望得到提高。


▲过量表达APLNR,让肿瘤细胞溶解的更多(图片来源:《自然》)

总结来说,我们用CRISPR-Cas9基因编辑技术,筛选出了一个关键的基因APLNR。它的突变会导致免疫疗法的效果下降。科学家们相信,这不但为我们提供了一个潜在增强免疫疗法的新靶点,还表明CRISPR-Cas9系统有望能用于寻找个体化的基因突变,加速个体化的新药开发。

也许不久后,每名癌症患者都能迎来最适合自己,也最有效的免疫疗法。我们期待这一天尽早到来!
原始出处;
Glenn Merlino,Feng Zhang,Nicholas P. Restifo.et al.Identification of essential genes for cancer immunotherapy.Nature


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    2017-10-04 liye789132251
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    2017-08-09 yuandd
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    2017-08-08 蔬菜

    高大上的研究,好好学习啊

    0

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    2017-08-08 明月清辉

    谢谢分享,学习了

    0

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