整合素 αVβ3 拮抗剂-c(RGDyk) 肽通过抑制成骨和血管生成来减轻后纵韧带骨化的进展。
Integrin αVβ3 antagonist-c(RGDyk) peptide attenuates the progression of ossification of the posterior longitudinal ligament by inhibiting osteogenesis and angiogenesis.
机构信息
Spine Center, Department of Orthopaedics, Changzheng Hospital, Naval Medical University (Second Military Medical University), Shanghai, 200003, China.
出版信息
Mol Med. 2024 May 2;30(1):57. doi: 10.1186/s10020-024-00822-x.
BACKGROUND
Ossification of the posterior longitudinal ligament (OPLL), an emerging heterotopic ossification disease, causes spinal cord compression, resulting in motor and sensory dysfunction. The etiology of OPLL remains unclear but may involve integrin αVβ3 regulating the process of osteogenesis and angiogenesis. In this study, we focused on the role of integrin αVβ3 in OPLL and explored the underlying mechanism by which the c(RGDyk) peptide acts as a potent and selective integrin αVβ3 inhibitor to inhibit osteogenesis and angiogenesis in OPLL.
METHODS
OPLL or control ligament samples were collected in surgery. For OPLL samples, RNA-sequencing results revealed activation of the integrin family, particularly integrin αVβ3. Integrin αVβ3 expression was detected by qPCR, Western blotting, and immunohistochemical analysis. Fluorescence microscopy was used to observe the targeted inhibition of integrin αVβ3 by the c(RGDyk) peptide on ligaments fibroblasts (LFs) derived from patients with OPLL and endothelial cells (ECs). The effect of c(RGDyk) peptide on the ossification of pathogenic LFs was detected using qPCR, Western blotting. Alkaline phosphatase staining or alizarin red staining were used to test the osteogenic capability. The effect of the c(RGDyk) peptide on angiogenesis was determined by EC migration and tube formation assays. The effects of the c(RGDyk) peptide on heterotopic bone formation were evaluated by micro-CT, histological, immunohistochemical, and immunofluorescence analysis in vivo.
RESULTS
The results indicated that after being treated with c(RGDyk), the osteogenic differentiation of LFs was significantly decreased. Moreover, the c(RGDyk) peptide inhibited the migration of ECs and thus prevented the nutritional support required for osteogenesis. Furthermore, the c(RGDyk) peptide inhibited ectopic bone formation in mice. Mechanistic analysis revealed that c(RGDyk) peptide could inhibit osteogenesis and angiogenesis in OPLL by targeting integrin αVβ3 and regulating the FAK/ERK pathway.
CONCLUSIONS
Therefore, the integrin αVβ3 appears to be an emerging therapeutic target for OPLL, and the c(RGDyk) peptide has dual inhibitory effects that may be valuable for the new therapeutic strategy of OPLL.
背景
骨化性后纵韧带（OPLL）是一种新兴的异位骨化疾病，可导致脊髓受压，引起运动和感觉功能障碍。OPLL 的病因尚不清楚，但可能涉及整合素 αVβ3 调节成骨和血管生成过程。在这项研究中，我们专注于整合素 αVβ3 在 OPLL 中的作用，并探讨了 c（RGDyk）肽作为一种有效的、选择性整合素 αVβ3 抑制剂，抑制 OPLL 中成骨和血管生成的潜在机制。
方法
在手术中收集 OPLL 或对照韧带样本。对于 OPLL 样本，RNA 测序结果显示整合素家族的激活，特别是整合素 αVβ3。通过 qPCR、Western blot 和免疫组织化学分析检测整合素 αVβ3 的表达。荧光显微镜用于观察靶向抑制患者 OPLL 韧带成纤维细胞（LFs）和内皮细胞（ECs）中整合素 αVβ3 的 c（RGDyk）肽。通过 qPCR、Western blot 检测 c（RGDyk）肽对致病 LFs 骨化的影响。碱性磷酸酶染色或茜素红染色用于检测成骨能力。通过 EC 迁移和管形成试验测定 c（RGDyk）肽对血管生成的影响。通过体内 micro-CT、组织学、免疫组织化学和免疫荧光分析评价 c（RGDyk）肽对异位骨形成的影响。
结果
结果表明，用 c（RGDyk）处理后，LFs 的成骨分化明显降低。此外，c（RGDyk）肽抑制 EC 的迁移，从而阻止成骨所需的营养支持。此外，c（RGDyk）肽抑制小鼠异位骨形成。机制分析表明，c（RGDyk）肽通过靶向整合素 αVβ3 并调节 FAK/ERK 通路，抑制 OPLL 中的成骨和血管生成。
结论
因此，整合素 αVβ3 似乎是 OPLL 的一个新兴治疗靶点，c（RGDyk）肽具有双重抑制作用，可能对 OPLL 的新治疗策略具有价值。
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