PO.CL05.03 · 临床研究
CREPT-618：首个靶向CREPT的GalNAc-siRNA逆转HCC免疫逃逸并与抗PD-1协同
Crept-618: First GalNAc-siRNA targeting CREPT reverses HCC immune evasion and synergizes with anti-PD-1
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
目的：免疫检查点抑制剂（ICIs）由于肿瘤呈免疫学"冷"表型（其特征为T细胞浸润差），仅在约~20%的肝细胞癌（HCC）患者中获得应答。我们确认CREPT在HCC中过表达，并与更差的生存（HR=2.722，P=0.0018）和ICI无应答相关（无应答者中表达更高；GSE215011，P<0.01）——它是肿瘤增殖和免疫沉默的双重驱动因素。我们提出CREPT-618，首个靶向CREPT的GalNAc偶联siRNA。
实验流程：在TCGA、GEO、CPTAC数据库中分析了CREPT表达，并在一个104例患者的HCC组织微阵列中通过免疫组织化学进行验证。通过在HCC细胞系（LM3、Huh7、Hep3B和Hepa1-6）中敲低CREPT，采用增殖、迁移和侵袭实验检测其功能作用。在有或无抗PD-1联合的人源化原位植入小鼠模型中评估体内疗效。使用RNA-seq、ATAC-seq、ChIP-seq和scRNA-seq研究其潜在机制。在小鼠、大鼠和猴中检测了CREPT-618的效力（无转染）、特异性、PK、生物分布（Cy5/LC-MS）以及GLP毒理学。
新的未发表数据：CREPT将SIN3A-HDAC1/2复合物募集至CCL5启动子，导致染色质压缩（ATAC-seq）并沉默IFN-gamma/IRF1诱导的CCL5转录（ChIP-seq）。CREPT缺失增加了CCL5分泌（3.7倍，P<0.001），驱动CD8+ T/NK细胞浸润（2.8倍，P<0.01；scRNA-seq）——该效应可被CCR5阻断所阻止，证实了对CCL5的依赖性。CREPT-618实现了强效、特异性沉默（IC50 0.046-1.628 nM；经RNA-seq/PCA验证），具有肝脏聚焦的PK特征（Cmax ~200 μg/g，T½ ~70 h）。在原位HCC中，单药治疗在3个批次中适度抑制了生长（IHC确认靶点结合），而与抗PD-1联合驱动了协同性的78.3%肿瘤消退（P<0.001）和细胞毒性CD8+ T细胞5.1倍的增加。CREPT-618在啮齿动物中高达100 mg/kg、在猴中高达30 mg/kg时耐受良好。
结论：CREPT通过抑制CCL5、经由CCL5-CCR5轴阻断T细胞募集，从而驱动HCC生长和免疫逃逸。CREPT-618——首个靶向CREPT的siRNA——沉默这一致癌枢纽，恢复CCL5信号，将"冷"肿瘤转化为"热"肿瘤，并与抗PD-1协同。这些临床前数据将支持一项首次人体试验；初步的人体安全性和CREPT敲低数据将在AACR上公布。
查看英文原文 English abstract
Purpose: Immune checkpoint inhibitors (ICIs) achieve responses in only approximately ~20% of hepatocellular carcinoma (HCC) patients due to immunologically “cold” tumors characterized by poor T-cell infiltration. We identify CREPT-overexpressed in HCC and linked to worse survival (HR=2.722, P=0.0018) and ICI non-response (higher expression in non-responders; GSE215011, P<0.01)-as a dual driver of tumor proliferation and immune silencing. We present CREPT-618, the first GalNAc-conjugated siRNA targeting CREPT.
Experimental Procedures: CREPT expression was analyzed in TCGA, GEO, CPTAC databases and validated by immunohistochemistry in a 104-patient HCC tissue microarray. Functional roles were tested via CREPT knockdown in HCC cell lines (LM3, Huh7, Hep3B and Hepa1-6) using proliferation, migration, and invasion assays. In vivo efficacy was assessed in humanized orthotopic implantation mouse models with or without anti-PD-1 combination. The underlying mechanism was studied using RNA-seq, ATAC-seq, ChIP-seq, and scRNA-seq. CREPT-618 was tested for potency (no transfection), specificity, PK, biodistribution (Cy5/LC-MS), and GLP toxicology in mice, rats, and monkeys.
New, Unpublished Data: CREPT recruits the SIN3A-HDAC1/2 complex to the CCL5 promoter, leading to chromatin compaction (ATAC-seq) and silencing IFN-gamma/IRF1-induced CCL5 transcription (ChIP-seq). CREPT loss increased CCL5 secretion (3.7-fold, P<0.001), driving CD8+ T/NK infiltration (2.8-fold, P<0.01; scRNA-seq)-prevented by CCR5 blockade, confirming CCL5 dependence. CREPT-618 achieved potent, specific silencing (IC₅₀ 0.046-1.628 nM; RNA-seq/PCA validated) with liver-focused PK (Cmax ~200 μg/g, T½ ~70 h). In orthotopic HCC, monotherapy modestly suppressed growth across 3 batches (IHC target engagement confirmed), while combination with anti-PD-1 drove synergistic 78.3% tumor regression (P<0.001) and a 5.1-fold increase in cytotoxic CD8+ T cells. CREPT-618 was well-tolerated up to 100 mg/kg (rodents) and 30 mg/kg (monkeys).
Conclusion: CREPT drives HCC growth and immune evasion by repressing CCL5, blocking T-cell recruitment via the CCL5-CCR5 axis. CREPT-618-the first siRNA targeting CREPT-silences this oncogenic hub, restores CCL5 signaling, converts “cold” tumors to “hot”, and synergizes with anti-PD-1. These preclinical data will enable a first-in-human trial; preliminary human safety and CREPT knockdown data will be presented at AACR.
利益披露 Disclosure
J. Li,
Heya Therapeutics ).
A. Zou, None.
H. Yang,
Heya Therapuetics ).
J. Wang,
Heya Therapeutics ).
W. Yang,
Heya Therapuetics ).
Z. Chang,
Heya Therapuetics ).
J. Li,
Heya Therapeutics ).