PO.TB10.12 · 肿瘤生物学
基质硬度诱导Ca²⁺-DCLK1-PIP5K1A机械转导，作为胰腺癌进展和化疗耐药中的一个生物力学检查点
Matrix stiffness induces Ca²⁺-DCLK1-PIP5K1A mechanotransduction as a biomechanical checkpoint in pancreatic cancer progression and chemotherapy resistance
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摘要 Abstract
中文摘要
背景：细胞外基质（ECM）结构和硬度的改变是胰腺癌快速进展的标志。然而，ECM生物力学特性影响恶性生物学行为的机制在很大程度上仍不清楚。钙调蛋白依赖性蛋白激酶DCLK1已被认为与癌症进展相关，但其在整合胰腺癌生物力学信号中的作用尚未阐明。
方法：我们使用体外生物力学应激模型和体内实体瘤实验，研究了胰腺癌中ECM硬度与DCLK1激活之间的关系。通过过表达或敲低来操控DCLK1的表达和活性，并使用特异性抑制剂调节钙信号。进行单细胞RNA测序以识别钙抑制使肿瘤对化疗增敏的潜在通路。对临床肿瘤样本进行多色免疫荧光染色，以原位检查PIEZO1-DCLK1-PIP5K1A-AKT信号轴与ECM硬度之间的相关性。机制研究包括蛋白相互作用检测和磷酸化分析，以界定DCLK1-PIP5K1A-PI3K-AKT信号级联。
结果：DCLK1的表达和激活在高生物力学应激下经PIEZO1/钙/HPCAL1轴介导被选择性诱导。在低硬度条件下过表达DCLK1加速了肿瘤进展和化疗耐药，而这一效应可被钙抑制剂部分逆转。相反，在高硬度条件下，敲低DCLK1抑制了肿瘤生长并增加了化疗敏感性，但削弱了联合钙抑制剂治疗的增敏效应。单细胞RNA测序识别出参与化疗增敏的钙相关通路。在机制上，DCLK1通过抑制PIP5K1A的苏氨酸磷酸化与其相互作用，促进PIP5K1A的膜定位并激活下游PI3K-AKT通路。多色免疫荧光在临床样本中证实了PIEZO1-DCLK1-PIP5K1A-AKT激活与ECM硬度的相关性。
结论：DCLK1作为胰腺癌中的生物力学检查点，整合ECM来源的机械线索以加剧肿瘤进展和化疗耐药。靶向钙/DCLK1信号轴可能增强胰腺癌患者辅助治疗的疗效。
查看英文原文 English abstract
Background: Alterations in extracellular matrix (ECM) architecture and stiffness are hallmarks of rapid pancreatic cancer progression. However, the mechanisms by which ECM biomechanical properties influence malignant biological behavior remain largely unknown. Calmodulin-dependent protein kinase DCLK1 has been implicated in cancer progression, but its role in integrating biomechanical signals in pancreatic cancer has not been elucidated.
Methods: We investigated the relationship between ECM stiffness and DCLK1 activation in pancreatic cancer using in vitro biomechanical stress models and in vivo solid tumor experiments. DCLK1 expression and activity were manipulated via overexpression or knockdown, and calcium signaling was modulated using specific inhibitors. Single-cell RNA sequencing was performed to identify potential pathways by which calcium inhibition sensitizes tumors to chemotherapy. Multicolor immunofluorescence staining of clinical tumor samples was used to examine the correlation between the PIEZO1-DCLK1-PIP5K1A-AKT signaling axis and ECM stiffness in situ. Mechanistic studies included protein interaction assays and phosphorylation analyses to define the DCLK1-PIP5K1A-PI3K-AKT signaling cascade.
Results: DCLK1 expression and activation were selectively induced under high biomechanical stress mediated by the PIEZO1/calcium/HPCAL1 axis. Overexpression of DCLK1 under low stiffness conditions accelerated tumor progression and chemoresistance, which could be partially reversed by calcium inhibitors. Conversely, under high stiffness conditions, DCLK1 knockdown inhibited tumor growth and increased chemosensitivity, but attenuated the sensitizing effect of combined calcium inhibitor treatment. Single-cell RNA sequencing identified calcium-related pathways contributing to chemotherapy sensitization. Mechanistically, DCLK1 interacted with PIP5K1A by inhibiting its threonine phosphorylation, promoting membrane localization of PIP5K1A and activating the downstream PI3K-AKT pathway. Multicolor immunofluorescence confirmed the correlation of PIEZO1-DCLK1-PIP5K1A-AKT activation with ECM stiffness in clinical samples.
Conclusions: DCLK1 functions as a biomechanical checkpoint in pancreatic cancer, integrating ECM-derived mechanical cues to exacerbate tumor progression and chemotherapy resistance. Targeting the calcium/DCLK1 signaling axis may enhance the efficacy of adjuvant therapy in pancreatic cancer patients
利益披露 Disclosure
H. Zhang, None..
C. Zhao, None..
J. Chen, None..
X. Hu, None..
J. Bai, None..
L. He, None..
Z. Deng, None..
T. Yin, None.