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重组表达一种新型纤维连接蛋白-胶原融合肽，通过整合素β3调节干细胞干性。

Recombinant expression a novel fibronectin-collage fusion peptide modulating stem cell stemness via integrin β3.

机构信息

Institute of Biomedicine and Guangdong Provincial Key Laboratory of Bioengineering Medicine, Jinan University, Guangzhou, 510632, China.

Biopharmaceutical R&D Center of Jinan University, Guangzhou, China.

出版信息

Appl Microbiol Biotechnol. 2022 May;106(9-10):3765-3776. doi: 10.1007/s00253-022-11965-4. Epub 2022 May 20.

DOI:10.1007/s00253-022-11965-4
PMID:35590080
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9151557/
Abstract

Constructing bionic extracellular matrix (ECM) is an attractive proposition for tissue engineering and clinical regeneration therapy involving the stemness of stem cells. Here, a novel recombinant protein fibronectin-collagen peptide (FCP) was designed to modulate the function of ECM expressed by Picha. pastoris strain X33. This FCP promotes cell migration and adhesion and maintains rBMSC stemness by binding integrin β3. Its effects were blocked by both integrin β3 siRNA and the integrin β3 inhibitor Cilengitide. A template-independent ab initio prediction modeling approach is the best approach to construct a stable FCP protein model, which predicts the binding sites between FCP and integrin β3. FCP may be used in the in vitro culture and clinical regeneration of stem cells that highly express integrin β3, such as hematopoietic stem cells. The study provides information on the molecular structure of FCP and its bioactivity, which can be used to design new compounds. KEY POINTS: • Design a novel recombinant fibronectin-collagen peptide biomimetic ECM. • FCP promotes cell adhesion, migration, and proliferation. • Predicted and verified FCP structure and affinity with integrin β3. • FCP binds integrin β3 to maintain rBMSC stemness.

摘要

构建仿生细胞外基质 (ECM) 是组织工程和涉及干细胞干性的临床再生治疗的一个有吸引力的方案。在这里，设计了一种新型重组蛋白纤维连接蛋白-胶原肽 (FCP) 来调节毕赤酵母菌株 X33 表达的 ECM 的功能。这种 FCP 通过结合整合素 β3 促进细胞迁移和黏附，并维持 rBMSC 的干性。其作用被整合素 β3 siRNA 和整合素 β3 抑制剂 cilengitide 阻断。无模板从头预测建模方法是构建稳定 FCP 蛋白模型的最佳方法，该方法预测了 FCP 和整合素 β3 之间的结合位点。FCP 可用于高度表达整合素 β3（如造血干细胞）的干细胞的体外培养和临床再生。该研究提供了关于 FCP 的分子结构和生物活性的信息，可用于设计新的化合物。

关键点

  1. 设计新型重组纤维连接蛋白-胶原肽仿生 ECM。

  2. FCP 促进细胞黏附、迁移和增殖。

  3. 预测和验证了 FCP 结构及其与整合素 β3 的亲和力。

  4. FCP 通过结合整合素 β3 维持 rBMSC 的干性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/82c7ef2b7164/253_2022_11965_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/3cd12139b979/253_2022_11965_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/668cc23d9f52/253_2022_11965_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/b92ff30d839a/253_2022_11965_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/6e99f5c582c0/253_2022_11965_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/82c7ef2b7164/253_2022_11965_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/3cd12139b979/253_2022_11965_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/668cc23d9f52/253_2022_11965_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/b92ff30d839a/253_2022_11965_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/6e99f5c582c0/253_2022_11965_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7368/9151557/82c7ef2b7164/253_2022_11965_Fig5_HTML.jpg

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