Druzhkova Irina N, Orlova Anna G, Fedulova Anastasiia S, Avakiants Arina V, Isakova Alina A, Kukovyakina Ekaterina V, Zijian Yuan, Plotnikova Ekaterina A, Trunova Galina V, Pankratov Andrey A, Plekhanov Anton A, Kurnikov Alexey A, Subochev Pavel V, Shaytan Alexey K, Gasparian Marine E, Kirpichnikov Mikhail P, Dolgikh Dmitry A, Razansky Daniel, Yagolovich Anne V
Privolzhsky Research Medical University, Nizhny Novgorod, 603081, Russia.
A.V. Gaponov-Grekhov Institute of Applied Physics of the Russian Academy of Sciences, Nizhny Novgorod, 603950, Russia.
J Transl Med. 2025 Aug 21;23(1):949. doi: 10.1186/s12967-025-06859-8.
Anti-angiogenic therapy is a clinically validated method for cancer treatment. It was previously revealed that concurrent targeting of angiogenic and death receptor signaling pathways by a multivalent DR5-specific cytokine TRAIL variant DR5-B genetically fused with the effector peptides, SRH-DR5-B-iRGD, enhances solid tumor suppression and prolongs survival. The SRH peptide is aimed at blocking the tumor neoangiogenesis by preventing activation of the VEGFR2 receptor, while the iRGD peptide interferes with the activation of integrin αβ, and enhances the tumor penetration. Here, we investigated how the antiangiogenic activity of the SRH-DR5-B-iRGD fusion protein contributes to its antitumor effects.
An integrated approach has been applied involving molecular modeling of SRH-DR5-B-iRGD binding to DR5 receptor, optoacoustic (OA) and optical coherence tomography-based microangiography (OCT-MA) imaging of the vessel networks in xenografts of human glioblastoma and pancreatic adenocarcinoma in nude mice, supported by immunohistochemical (IHC) staining for vascularization marker CD31, and in vitro and in vivo bioactivity studies.
Molecular modeling has demonstrated that genetic fusion of DR5-B with the SRH and iRGD peptides not only enables the engagement of additional tumor targets VEGFR2 and integrin αβ/NRP-1, but also improves the interaction with DR5 receptor. OA imaging of the vessel network in xenograft tumor nodes of human glioblastoma and pancreatic adenocarcinoma displayed a decrease in the vessel fraction in DR5-B-treated xenograft tumors, with the effect being even more pronounced in SRH-DR5-B-iRGD-treated tumor nodes. This data was consistent with the reduction in the number of perfused vessels in DR5-B and SRH-DR5-B-iRGD-treated tumors as quantified by OCT-MA, and also correlate well with the data obtained by IHC staining and tumor growth inhibition.
Ameliorated interaction with the DR5 receptor and imparting antiangiogenic properties to the multivalent fusion protein SRH-DR5-B-iRGD resulted in improved antitumor activity compared to DR5-B. Thereby, SRH-DR5-B-iRGD can be considered as a promising candidate for the treatment of vascularized solid tumors.
抗血管生成疗法是一种经过临床验证的癌症治疗方法。先前的研究表明，一种与效应肽基因融合的多价DR5特异性细胞因子TRAIL变体DR5-B（SRH-DR5-B-iRGD）同时靶向血管生成和死亡受体信号通路，可增强实体瘤抑制并延长生存期。SRH肽旨在通过阻止VEGFR2受体的激活来阻断肿瘤新生血管生成，而iRGD肽则干扰整合素αβ的激活，并增强肿瘤渗透。在此，我们研究了SRH-DR5-B-iRGD融合蛋白的抗血管生成活性如何对其抗肿瘤作用产生影响。
采用了一种综合方法，包括对SRH-DR5-B-iRGD与DR5受体结合的分子建模、基于光声（OA）和光学相干断层扫描的微血管造影（OCT-MA）对裸鼠人胶质母细胞瘤和胰腺腺癌异种移植瘤中的血管网络进行成像，并辅以血管生成标记物CD31的免疫组织化学（IHC）染色，以及体外和体内生物活性研究。
分子建模表明，DR5-B与SRH和iRGD肽的基因融合不仅能够结合额外的肿瘤靶点VEGFR2和整合素αβ/NRP-1，还能改善与DR5受体的相互作用。对人胶质母细胞瘤和胰腺腺癌异种移植瘤结节中的血管网络进行OA成像显示，DR5-B处理的异种移植瘤中血管分数降低，而在SRH-DR5-B-iRGD处理的肿瘤结节中这种效果更为明显。该数据与通过OCT-MA定量的DR5-B和SRH-DR5-B-iRGD处理的肿瘤中灌注血管数量的减少一致，并且也与IHC染色和肿瘤生长抑制获得的数据高度相关。
与DR5-B相比，SRH-DR5-B-iRGD与DR5受体的相互作用得到改善，并赋予多价融合蛋白抗血管生成特性，从而导致抗肿瘤活性增强。因此，SRH-DR5-B-iRGD可被视为治疗血管化实体瘤的有前景的候选药物。
