CLK1 通过CK2α激活HIPPO信号通路对肝再生的影响及机制研究

Liver regeneration plays a crucial role in the recovery of hepatectomy and liver function after injury, and its molecular mechanism is more complex. CLK1 is a bispecific Serine/threonine and tyrosine protein kinase that regulates cell proliferation, differentiation, and pre-mRNA splicing. However, it is unclear whether CLK1 is involved in regulating liver regeneration. Previously, we found that knocking down CLK1 in mouse liver can significantly inhibit the activity of HIPPO signaling pathway and promote regeneration of residual liver after 2/3 hepatectomy in mice. Subsequent screenings revealed that CLK1 interacted with CK2α. On this basis, we intend to observe the liver regeneration and HIPPO pathway activity changes by intervening CLK1 and CK2α in mouse liver. CLK1 inhibitors were used to observe their effects on liver regeneration in mice, and the potential of CLK1 as a therapeutic target for liver regeneration was explored. We aim to examin the molecular mechanism of CLK1 regulation of HIPPO pathway activity through CK2α, to clarify the role and mechanism of CLK1/CK2α/HIPPO pathway in liver regeneration, which providing a theoretical support for further study of the molecular mechanisms regulating liver regeneration, and for discovery and screening new treatment drugs of liver regeneration.

肝再生对行肝切除术及损伤后肝功能的恢复起到至关重要的作用，其分子机制较为复杂。CLK1是一种双特异性丝/苏氨酸和酪氨酸蛋白激酶，具有调节细胞增殖、分化及RNA剪接等作用。然而，CLK1是否参与调节肝再生尚不清楚。前期我们发现，敲低小鼠肝脏中CLK1可明显抑制HIPPO信号通路活性，促进小鼠2/3肝切除术后的残肝再生。后续通过筛选发现CLK1与CK2α具有相互作用。在此基础上，我们拟通过干预小鼠肝脏中的CLK1和CK2α，观察其肝再生的情况及HIPPO通路活性变化。采用CLK1抑制剂观察其对小鼠肝再生的影响，初步探讨CLK1作为肝再生治疗靶点的潜力。本研究旨在明确CLK1通过CK2α调控HIPPO通路活性的分子机制，阐明CLK1/CK2α/HIPPO通路在肝再生中的作用及机制，为深入研究调控肝再生的分子机制奠定理论基础，为发现和筛选新的促肝再生治疗药物提供理论依据。

肝再生对行肝切除术及损伤后肝功能的恢复起到至关重要的作用，其分子机制较为复杂。CLK1是一种双特异性丝/苏氨酸和酪氨酸蛋白激酶，具有调节细胞增殖、分化及RNA剪接等作用。然而，CLK1是否参与调节肝再生尚不清楚。本研究分别采用干扰和过表达小鼠肝脏中CLK1后，建立小鼠2/3肝切术模型和CCL4肝损伤模型，证实了CLK1对肝再生过程中的影响，进一步采用生物大分子相互作用检测和co-IP验证明确了CLK1能够通过CK2α/HIPPO通路影响肝再生过程。细胞水平上，使用AgILent细胞无标记靶向药物分析仪，实时、动态、连续检测小鼠正常肝脏细胞CLK1稳定干扰株和稳定过表达株的细胞增殖情况，得到了和体内一致的结果。进一步发现，在一定浓度范围内，CLK1抑制剂能够促进正常肝细胞增殖和抑制肝癌细胞增殖，提示CLK1在肝再生中是一个较为理想的靶点。综上，本研究阐明了CLK1通过CK2α调控HIPPO通路在肝再生中的作用机制，为深入研究调控肝再生的分子机制奠定理论基础，为发现和筛选新的促肝再生治疗药物提供新的靶点。