Zaninelli Tiago H, Fattori Victor, Heintz Olivia K, Wright Kristeena R, Bennallack Philip R, Sim Danielle, Bukhari Hussain, Terry Kathryn L, Vitonis Allison F, Missmer Stacey A, Andrello Avacir C, Anchan Raymond M, Godin Stephen K, Bree Dara, Verri Waldiceu A, Rogers Michael S
Vascular Biology Program, Department of Surgery, Boston Children's Hospital, Harvard Medical School, Boston, MA, United States; Laboratory of Pain, Inflammation, Neuropathy, and Cancer, Department of Pathology, Center of Biological Sciences, Londrina State University, Londrina, PR, Brazil.
Vascular Biology Program, Department of Surgery, Boston Children's Hospital, Harvard Medical School, Boston, MA, United States.
J Adv Res. 2025 Jul;73:593-605. doi: 10.1016/j.jare.2024.08.017. Epub 2024 Aug 12.
Endometriosis is a chronic inflammatory disease that affects ∼10 % of women. A significant fraction of patients experience limited or no efficacy with current therapies. Tissue adjacent to endometriosis lesions often exhibits increased neurite and vascular density, suggesting that disease pathology involves neurotrophic activity and angiogenesis.
We aim to evaluate the potential for key tyrosine-kinase-receptor-coupled neurotrophic molecules to contribute to endometriosis-associated pain in mice.
Peritoneal fluid was collected from endometriosis patients undergoing surgery and the levels of NGF and VEGFR1 regulators (VEGFA, VEGFB, PLGF, and sVEGFR1) were quantified by ELISA. VEGFR1 regulator concentrations were used to calculate VEGFR1 occupancy. We used genetic depletion, neutralizing antibodies, and pharmacological approaches to specifically block neurotrophic ligands (NGF or BDNF) or receptors (VEGFR1, TRKs) in a murine model of endometriosis-associated pain. Endometriosis-associated pain was measured using von Frey filaments, quantification of spontaneous abdominal pain-related behavior, and thermal discomfort. Disease parameters were evaluated by lesion size and prevalence. To evaluate potential toxicity, we measured the effect of entrectinib dose and schedule on body weight, liver and kidney function, and bone structure (via micro-CT).
We found that entrectinib (pan-Trk inhibitor) or anti-NGF treatments reduced evoked pain, spontaneous pain, and thermal discomfort. In contrast, even though calculated receptor occupancy revealed that VEGFR1 agonist levels are sufficient to support signaling, blocking VEGFR1 via antibody or tamoxifen-induced knockout did not reduce pain or lesion size in mice. Targeting BDNF-TrkB with an anti-BDNF antibody also proved ineffective. Notably, changing dosing schedule to once weekly eliminated entrectinib-induced bone-loss without decreasing efficacy against pain.
This suggests NGF-TrkA signaling, but not BDNF-TrkB or VEGF-VEGFR1, mediates endometriosis-associated pain. Moreover, entrectinib blocks endometriosis-associated pain and reduces lesion sizes. Our results also indicated that entrectinib-like molecules are promising candidates for endometriosis treatment.
子宫内膜异位症是一种慢性炎症性疾病，影响约10%的女性。相当一部分患者对当前治疗的疗效有限或无效。子宫内膜异位症病灶周围的组织通常表现出神经突和血管密度增加，这表明疾病病理涉及神经营养活性和血管生成。
我们旨在评估关键的酪氨酸激酶受体偶联神经营养分子对小鼠子宫内膜异位症相关疼痛的影响。
收集接受手术的子宫内膜异位症患者的腹腔液，通过酶联免疫吸附测定法（ELISA）定量神经生长因子（NGF）和血管内皮生长因子受体1（VEGFR1）调节剂（血管内皮生长因子A [VEGFA]、血管内皮生长因子B [VEGFB]、胎盘生长因子 [PLGF] 和可溶性血管内皮生长因子受体1 [sVEGFR1]）的水平。VEGFR1调节剂浓度用于计算VEGFR1占有率。我们使用基因敲除、中和抗体和药理学方法在子宫内膜异位症相关疼痛的小鼠模型中特异性阻断神经营养配体（NGF或脑源性神经营养因子 [BDNF]）或受体（VEGFR1、酪氨酸激酶受体 [TRKs]）。使用von Frey细丝、定量自发性腹痛相关行为和热不适来测量子宫内膜异位症相关疼痛。通过病灶大小和患病率评估疾病参数。为了评估潜在毒性，我们测量了恩曲替尼剂量和给药方案对体重、肝肾功能和骨结构（通过微型计算机断层扫描 [micro-CT]）的影响。
我们发现恩曲替尼（泛TRK抑制剂）或抗NGF治疗可减轻诱发性疼痛、自发性疼痛和热不适。相比之下，尽管计算出的受体占有率显示VEGFR1激动剂水平足以支持信号传导，但通过抗体或他莫昔芬诱导的基因敲除阻断VEGFR1并不能减轻小鼠的疼痛或病灶大小。用抗BDNF抗体靶向BDNF-TrkB也被证明无效。值得注意的是，将给药方案改为每周一次可消除恩曲替尼引起的骨质流失，而不会降低对疼痛的疗效。
这表明NGF-TrkA信号传导而非BDNF-TrkB或血管内皮生长因子-血管内皮生长因子受体1（VEGF-VEGFR1）介导子宫内膜异位症相关疼痛。此外，恩曲替尼可阻断子宫内膜异位症相关疼痛并减小病灶大小。我们的结果还表明，类似恩曲替尼的分子是子宫内膜异位症治疗的有希望的候选药物。
