Lee Jun Hee, Kim Sung Wook, Ji Seung Taek, Kim Yeon Ju, Jang Woong Bi, Oh Jin-Woo, Kim Jaeho, Yoo So Young, Beak Sang Hong, Kwon Sang-Mo
1Department of Pharmacology and Toxicology, University of Alabama at Birmingham School of Medicine, Birmingham, AL 35294 USA.
2Laboratory for Vascular Medicine and Stem Cell Biology, Department of Physiology, School of Medicine, Medical Research Institute, Pusan National University, 49, Busandaehak-ro, Mulgeum-eup, Yangsan, 50612 Republic of Korea.
Tissue Eng Regen Med. 2017 Aug 28;14(6):787-802. doi: 10.1007/s13770-017-0074-x. eCollection 2017 Dec.
Dysfunction or loss of blood vessel causes several ischemic diseases. Although endothelial progenitor cells (EPCs) are a promising source for cell-based therapy, ischemia-induced pathophysiological condition limits the recovery rate by causing drastic cell death. To overcome this issue, we attempted to develop a cell-targeted peptide delivery and priming system to enhance EPC-based neovascularization using an engineered M13 bacteriophage harboring nanofibrous tubes displaying ~2700 multiple functional motifs. The M13 nanofiber was modified by displaying RGD, which is an integrin-docking peptide, on the minor coat protein, and by mutilayering SDKP motifs, which are the key active sites for thymosin β4, on the major coat protein. The engineered M13 nanofiber dramatically enhanced ischemic neovascularization by activating intracellular and extracellular processes such as proliferation, migration, and tube formation in the EPCs. Furthermore, transplantation of the primed EPCs with the M13 nanofiber harboring RGD and SDKP facilitated functional recovery and neovascularization in a murine hindlimb ischemia model. Overall, this study demonstrates the effectiveness of the M13 nanofiber-based novel peptide delivery and priming strategy in promoting EPC bioactivity and neovessel regeneration. To our knowledge, this is first report on M13 nanofibers harboring dual functional motifs, the use of which might be a novel strategy for stem and progenitor cell therapy against cardiovascular ischemic diseases.
血管功能障碍或缺失会引发多种缺血性疾病。尽管内皮祖细胞(EPCs)是基于细胞治疗的一个有前景的细胞来源,但缺血诱导的病理生理状况会导致大量细胞死亡,从而限制了恢复率。为克服这一问题,我们尝试开发一种细胞靶向肽递送和预处理系统,利用携带展示约2700个多功能基序的纳米纤维管的工程化M13噬菌体来增强基于EPC的新生血管形成。通过在次要衣壳蛋白上展示整合素对接肽RGD,并在主要衣壳蛋白上多层展示胸腺素β4的关键活性位点SDKP基序,对M13纳米纤维进行了修饰。工程化的M13纳米纤维通过激活EPCs内的增殖、迁移和管形成等细胞内和细胞外过程,显著增强了缺血性新生血管形成。此外,用携带RGD和SDKP的M13纳米纤维预处理的EPCs移植,促进了小鼠后肢缺血模型中的功能恢复和新生血管形成。总体而言,本研究证明了基于M13纳米纤维的新型肽递送和预处理策略在促进EPC生物活性和新血管再生方面的有效性。据我们所知,这是关于携带双功能基序的M13纳米纤维的首次报道,其应用可能是针对心血管缺血性疾病的干细胞和祖细胞治疗的一种新策略。