Extra-renal expression of Cytochrome P450 Family 27 Subfamily B Member 1 (CYP27B1) has been well recognized and reflects the importance of intracrine/paracrine vitamin D signaling in different tissues under physiological and pathological conditions. In a prior RNA sequencing project, we identified CYP27B1 mRNA as upregulated in muscle samples from patients with amyotrophic lateral sclerosis (ALS) compared to normal controls. Our aims here were: (1) to validate this finding in a larger sample set including disease controls, (2) to determine which cell type is expressing CYP27B1 protein in muscle tissue, (3) to correlate CYP27B1 mRNA expression with disease progression in the SOD1G93A ALS mouse and in ALS patients. We assessed CYP27B1 expression by qPCR, western blot, and immunohistochemistry in a repository of muscle samples from ALS, disease controls (myopathy and non-ALS neuropathic disease), normal subjects, and muscle samples from the SOD1G93A mouse. Eight ALS patients were studied prospectively over 6-12 months with serial muscle biopsies. We found that CYP27B1 mRNA and protein levels were significantly increased in ALS versus normal and myopathy muscle samples. Neuropathy samples had increased CYP27B1 mRNA and protein expression but at a lower level than the ALS group. Immunohistochemistry showed that CYP27B1 localized to myofibers, especially those with features of denervation. In the SOD1G93A mouse, CYP27B1 mRNA and protein were detected in skeletal muscle in early pre-symptomatic stages and increased through end-stage. In the human study, increases in CYP27B1 mRNA in muscle biopsies correlated with disease progression rates over the same time period. In summary, we show for the first time that CYP27B1 mRNA and protein expression are elevated in muscle fibers in denervating disease, especially ALS, where mRNA levels can potentially serve as a surrogate marker for tracking disease progression. Its upregulation may reflect a local perturbation of vitamin D signaling, and further characterization of this pathway may provide insight into underlying molecular processes linked to muscle denervation.

中文翻译：

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维生素 D 激活剂 CYP27B1 在去神经疾病的肌纤维中表达上调，并且可以追踪肌萎缩侧索硬化症的进展。


细胞色素 P450 家族 27 亚家族 B 成员 1 (CYP27B1) 的肾外表达已得到充分认识，反映了生理和病理条件下不同组织中分泌/旁分泌维生素 D 信号传导的重要性。在之前的 RNA 测序项目中，我们发现与正常对照相比，肌萎缩侧索硬化症 (ALS) 患者的肌肉样本中 CYP27B1 mRNA 上调。我们的目标是：(1) 在包括疾病对照在内的更大样本集中验证这一发现，(2) 确定肌肉组织中表达 CYP27B1 蛋白的细胞类型，(3) 将 CYP27B1 mRNA 表达与肌肉组织中的疾病进展相关联。 SOD1G93A ALS 小鼠和 ALS 患者。我们通过 qPCR、蛋白质印迹和免疫组织化学方法评估了来自 ALS、疾病对照（肌病和非 ALS 神经性疾病）、正常受试者和 SOD1G93A 小鼠肌肉样本的肌肉样本库中 CYP27B1 的表达。通过连续肌肉活检对 8 名 ALS 患者进行了为期 6-12 个月的前瞻性研究。我们发现，与正常和肌病肌肉样本相比，ALS 中的 CYP27B1 mRNA 和蛋白质水平显着增加。神经病变样本的 CYP27B1 mRNA 和蛋白表达增加，但水平低于 ALS 组。免疫组织化学显示 CYP27B1 定位于肌纤维，尤其是具有去神经特征的肌纤维。在 SOD1G93A 小鼠中，在症状前早期阶段在骨骼肌中检测到 CYP27B1 mRNA 和蛋白质，并在晚期阶段增加。在人体研究中，肌肉活检中 CYP27B1 mRNA 的增加与同一时期的疾病进展率相关。 总之，我们首次表明，去神经疾病（尤其是 ALS）的肌纤维中 CYP27B1 mRNA 和蛋白表达升高，其中 mRNA 水平有可能作为跟踪疾病进展的替代标志物。它的上调可能反映了维生素 D 信号传导的局部扰动，对该通路的进一步表征可能有助于深入了解与肌肉去神经支配相关的潜在分子过程。