Targeting NGF but not VEGFR1 or BDNF signaling reduces endometriosis-associated pain in mice,Journal of Advanced Research

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Targeting NGF but not VEGFR1 or BDNF signaling reduces endometriosis-associated pain in mice
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2024-08-12 , DOI: 10.1016/j.jare.2024.08.017
Tiago H Zaninelli ₁ , Victor Fattori ₂ , Olivia K Heintz ₂ , Kristeena R Wright ₂ , Philip R Bennallack ₂ , Danielle Sim ₂ , Hussain Bukhari ₂ , Kathryn L Terry ₃ , Allison F Vitonis ₄ , Stacey A Missmer ₅ , Avacir C Andrello ₆ , Raymond M Anchan ₇ , Stephen K Godin ₈ , Dara Bree ₈ , Waldiceu A Verri ₉ , Michael S Rogers ₁₀

Affiliation

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.
Department of Obstetrics, Gynecology, and Reproductive Biology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, United States; Boston Center for Endometriosis, Boston Children's Hospital and Brigham and Women's Hospital, Boston, MA, United States; Department of Epidemiology, Harvard T.H. Chan School of Public Health, Boston, MA, United States.
Department of Obstetrics, Gynecology, and Reproductive Biology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, United States; Boston Center for Endometriosis, Boston Children's Hospital and Brigham and Women's Hospital, Boston, MA, United States.
Boston Center for Endometriosis, Boston Children's Hospital and Brigham and Women's Hospital, Boston, MA, United States; Department of Epidemiology, Harvard T.H. Chan School of Public Health, Boston, MA, United States; Department of Obstetrics, Gynecology, and Reproductive Biology, College of Human Medicine, Michigan State University, Grand Rapids, MI, United States.
Department of Physics, Center of Exact Sciences, Londrina State University, Londrina, PR, Brazil.
Boston Center for Endometriosis, Boston Children's Hospital and Brigham and Women's Hospital, Boston, MA, United States; Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, United States.
Cygnal Therapeutics, Cambridge, 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; Boston Center for Endometriosis, Boston Children's Hospital and Brigham and Women's Hospital, Boston, MA, United States.

Introduction

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.

Objectives

We aim to evaluate the potential for key tyrosine-kinase-receptor-coupled neurotrophic molecules to contribute to endometriosis-associated pain in mice.

Methods

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).

Results

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.

Conclusions

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.

中文翻译：

靶向 NGF 但不靶向 VEGFR1 或 BDNF 信号可减轻小鼠子宫内膜异位症相关疼痛

介绍

子宫内膜异位症是一种慢性炎症性疾病，影响约 10% 的女性。很大一部分患者对当前疗法的疗效有限或没有疗效。子宫内膜异位症病变附近的组织通常表现出神经突和血管密度增加，表明疾病病理涉及神经营养活动和血管生成。

目标

我们旨在评估关键酪氨酸激酶受体偶联神经营养分子导致小鼠子宫内膜异位症相关疼痛的潜力。

方法

收集子宫内膜异位症手术患者腹膜液，ELISA 定量 NGF 和 VEGFR1 调节因子（VEGFA、VEGFB、PLGF 和 sVEGFR1）水平。VEGFR1 调节因子浓度用于计算 VEGFR1 占有率。我们使用基因耗竭、中和抗体和药理学方法来特异性阻断子宫内膜异位症相关疼痛小鼠模型中的神经营养配体（NGF 或 BDNF）或受体（VEGFR1、TRKs）。使用 von Frey 细丝测量子宫内膜异位症相关疼痛，量化自发性腹痛相关行为和热不适。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时间表对体重、肝肾功能以及骨骼结构的影响（通过显微 CT）。

结果

我们发现恩曲替尼（pan-Trk 抑制剂）或抗 NGF 治疗减少了诱发痛、自发性疼痛和热不适。相比之下，即使计算的受体占有率显示 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敲除阻断 VEGFR1 并不能减轻小鼠的疼痛或病变大小。用抗 BDNF 抗体靶向 BDNF-TrkB 也被证明是无效的。值得注意的是，将给药方案更改为每周一次可消除恩曲替尼诱导的骨质流失，而不会降低对疼痛的疗效。