Wu Yiqian, Huang Ziliang, Liu Yahan, He Peixiang, Wang Yuxuan, Yan Liyanran, Wang Xinhui, Gao Shanzi, Zhou Xintao, Yoon Chi Woo, Sun Kun, Situ Yinglin, Ho Phuong, Zeng Yushun, Yuan Zhou, Zhu Linshan, Zhou Qifa, Zhao Yunde, Liu Thomas, Kwong Gabriel A, Chien Shu, Liu Longwei, Wang Yingxiao
Shu Chien - Gene Lay Department of Bioengineering, Institute of Engineering in Medicine, University of California San Diego, La Jolla, CA, USA.
National Biomedical Imaging Center, College of Future Technology, Peking University, Beijing, China.
Nat Commun. 2024 Dec 1;15(1):10444. doi: 10.1038/s41467-024-54477-7.
There remains a critical need for the precise control of CRISPR (clustered regularly interspaced short palindromic repeats)-based technologies. Here, we engineer a set of inducible CRISPR-based tools controllable by focused ultrasound (FUS), which can penetrate deep and induce localized hyperthermia for transgene activation. We demonstrate the capabilities of FUS-inducible CRISPR, CRISPR activation (CRISPRa), and CRISPR epigenetic editor (CRISPRee) in modulating the genome and epigenome. We show that FUS-CRISPR-mediated telomere disruption primes solid tumours for chimeric antigen receptor (CAR)-T cell therapy. We further deliver FUS-CRISPR in vivo using adeno-associated viruses (AAVs), followed by FUS-induced telomere disruption and the expression of a clinically validated antigen in a subpopulation of tumour cells, functioning as "training centers" to activate synthetic Notch (synNotch) CAR-T cells to produce CARs against a universal tumour antigen to exterminate neighboring tumour cells. The FUS-CRISPR(a/ee) toolbox hence allows the noninvasive and spatiotemporal control of genomic/epigenomic reprogramming for cancer treatment.
对于基于CRISPR（成簇规律间隔短回文重复序列）的技术，仍然迫切需要精确控制。在此，我们设计了一组可通过聚焦超声（FUS）控制的诱导型基于CRISPR的工具，其可穿透深部并诱导局部热疗以激活转基因。我们展示了FUS诱导型CRISPR、CRISPR激活（CRISPRa）和CRISPR表观遗传编辑器（CRISPRee）在调控基因组和表观基因组方面的能力。我们表明，FUS - CRISPR介导的端粒破坏使实体瘤对嵌合抗原受体（CAR）-T细胞疗法敏感。我们进一步使用腺相关病毒（AAV）在体内递送FUS - CRISPR，随后进行FUS诱导的端粒破坏以及在肿瘤细胞亚群中表达临床验证的抗原，作为“训练中心”来激活合成Notch（synNotch）CAR - T细胞，以产生针对通用肿瘤抗原的CAR来消灭邻近肿瘤细胞。因此，FUS - CRISPR（a/ee）工具箱允许对基因组/表观基因组重编程进行非侵入性的时空控制以用于癌症治疗。
