PDF(Pubmed)
Abstract：
BACKGROUND: Recent intravesical administration of adenoviral vectors, either as a single injection or in combination with immune checkpoint inhibitors, exemplified by cretostimogene grenadenorepvec and nadofaragene firadenovec, has demonstrated remarkable efficacy in clinical trials for non-muscle invasive bladder cancer. Despite their ability to induce an enhanced immune reaction within the lesion, the intracellular survival signaling of cancer cells has not been thoroughly addressed.
METHODS: An analysis of the prognostic data revealed a high probability of therapeutic efficacy with simultaneous inhibition of mTOR and STAT3. Considering the challenges of limited pharmaco-accessibility to the bladder due to its pathophysiological structure and the partially undruggable nature of target molecules, we designed a dual siRNA system targeting both mRNAs. Subsequently, this dual siRNA system was encoded into the adenovirus 5/3 (Ad 5/3) to enhance in vivo delivery efficiency.
RESULTS: Gene-targeting efficacy was assessed using cells isolated from xenografted tumors using a single-cell analysis system. Our strategy demonstrated a balanced downregulation of mTOR and STAT3 at the single-cell resolution, both in vitro and in vivo. This approach reduced tumor growth in bladder cancer xenograft and orthotopic animal experiments. In addition, increased infiltration of CD8+ T cells was observed in a humanized mouse model. We provided helpful and safe tissue distribution data for intravesical therapy of siRNAs coding adenoviruses.
CONCLUSIONS: The bi-specific siRNA strategy, encapsulated in an adenovirus, could be a promising tool to augment cancer treatment efficacy and overcome conventional therapy limitations associated with \"undruggability.\" Hence, we propose that dual targeting of mTOR and STAT3 is an advantageous strategy for intravesical therapy using adenoviruses.
METHODS: An analysis of the prognostic data revealed a high probability of therapeutic efficacy with simultaneous inhibition of mTOR and STAT3. Considering the challenges of limited pharmaco-accessibility to the bladder due to its pathophysiological structure and the partially undruggable nature of target molecules, we designed a dual siRNA system targeting both mRNAs. Subsequently, this dual siRNA system was encoded into the adenovirus 5/3 (Ad 5/3) to enhance in vivo delivery efficiency.
RESULTS: Gene-targeting efficacy was assessed using cells isolated from xenografted tumors using a single-cell analysis system. Our strategy demonstrated a balanced downregulation of mTOR and STAT3 at the single-cell resolution, both in vitro and in vivo. This approach reduced tumor growth in bladder cancer xenograft and orthotopic animal experiments. In addition, increased infiltration of CD8+ T cells was observed in a humanized mouse model. We provided helpful and safe tissue distribution data for intravesical therapy of siRNAs coding adenoviruses.
CONCLUSIONS: The bi-specific siRNA strategy, encapsulated in an adenovirus, could be a promising tool to augment cancer treatment efficacy and overcome conventional therapy limitations associated with \"undruggability.\" Hence, we propose that dual targeting of mTOR and STAT3 is an advantageous strategy for intravesical therapy using adenoviruses.
摘要：
背景：最近膀胱内给予腺病毒载体，作为单次注射或与免疫检查点抑制剂组合，以克构树基因grenadenorepvec和nadofaragenefiradenovec为例，在非肌层浸润性膀胱癌的临床试验中已经证明了显著的疗效。尽管它们能够在病变内诱导增强的免疫反应，癌细胞的细胞内存活信号尚未得到彻底解决。
方法：对预后数据的分析显示，同时抑制mTOR和STAT3的治疗效果的可能性很高。考虑到膀胱由于其病理生理结构和靶分子的部分不可药物性质而导致的有限的药物可及性的挑战，我们设计了一种靶向两种mRNA的双siRNA系统。随后，这种双重siRNA系统被编码到腺病毒5/3(Ad5/3)中以增强体内递送效率。
结果:使用单细胞分析系统,使用从异种移植肿瘤分离的细胞评估基因靶向功效。我们的策略展示了在单细胞分辨率下mTOR和STAT3的平衡下调，在体外和体内。这种方法减少了膀胱癌异种移植和原位动物实验中的肿瘤生长。此外,在人源化小鼠模型中观察到CD8+T细胞浸润增加。我们为编码腺病毒的siRNA的膀胱内治疗提供了有用且安全的组织分布数据。
结论：双特异性siRNA策略，封装在腺病毒中，可能是一个有希望的工具，以提高癌症治疗的疗效，并克服传统治疗的局限性，相关的\“不可药用。\"因此，我们认为mTOR和STAT3的双重靶向是使用腺病毒进行膀胱内治疗的有利策略.
方法：对预后数据的分析显示，同时抑制mTOR和STAT3的治疗效果的可能性很高。考虑到膀胱由于其病理生理结构和靶分子的部分不可药物性质而导致的有限的药物可及性的挑战，我们设计了一种靶向两种mRNA的双siRNA系统。随后，这种双重siRNA系统被编码到腺病毒5/3(Ad5/3)中以增强体内递送效率。
结果:使用单细胞分析系统,使用从异种移植肿瘤分离的细胞评估基因靶向功效。我们的策略展示了在单细胞分辨率下mTOR和STAT3的平衡下调，在体外和体内。这种方法减少了膀胱癌异种移植和原位动物实验中的肿瘤生长。此外,在人源化小鼠模型中观察到CD8+T细胞浸润增加。我们为编码腺病毒的siRNA的膀胱内治疗提供了有用且安全的组织分布数据。
结论：双特异性siRNA策略，封装在腺病毒中，可能是一个有希望的工具，以提高癌症治疗的疗效，并克服传统治疗的局限性，相关的\“不可药用。\"因此，我们认为mTOR和STAT3的双重靶向是使用腺病毒进行膀胱内治疗的有利策略.
