Abstract
Niemann-Pick disease type C1 (NPC1) is a hereditary neurodegenerative disorder caused by a mutation in the NPC1 gene. This gene encodes a transmembrane protein found in lysosomes. This disease characterized by hepatosplenomegaly, neurological impairments and premature death. Recent preclinical studies have shown promising results in using mesenchymal stem cells (MSCs) to alleviate the symptoms of NPC1. One type of MSCs, known as human menstrual blood-derived endometrial stem cells (MenSCs), has attracted attention due to its accessibility, abundant supply, and strong proliferation and regeneration capabilities. However, it remains uncertain whether the conditioned medium of MenSCs (MenSCs-CM) can effectively relieve the symptoms of NPC1. To investigate this further, we employed the CRISPR-Cas9 technique to successfully create a Npc1 gene knockout N2a cell line (Npc1KO N2a). Sanger sequencing confirmed the occurrence of Npc1 gene mutation in these cells, while western blotting revealed a lack of NPC1 protein expression. Filipin staining provided visual evidence of unesterified cholesterol accumulation in Npc1KO N2a cells. Moreover, Npc1KO N2a cells exhibited significantly decreased viability, increased inflammation, and heightened cell apoptosis. Notably, our study demonstrated that the viability of Npc1KO N2a cells was most significantly improved after being cultured by 36 h-collected MenSCs-CM for 0.5 days. Additionally, MenSCs-CM exhibited the ability to effectively reduce inflammation, counteract cell apoptosis, and ameliorate unesterified cholesterol accumulation in Npc1KO N2a cells. This groundbreaking finding establishes, for the first time, the protective effect of MenSCs-CM on N2a cells with Npc1 gene deletion. These findings suggest that the potential of MenSCs-CM as a beneficial therapeutic approach for NPC1 and other neurodegenerative diseases.
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We sincerely thank Dr. Chao Tong (University of Pennsylvania) for embellishing the language of the article.
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This work was supported by grants from the National Natural Science Foundation of China (grant number 81801127), Natural Science Foundation of Henan Province for Distinguished Young Scholars (grant number 202300410307), Central Guidance on Local Science and Technology Development Fund of Henan Province, Key Scientific Research Program of Higher Education in Henan Province (grant number 23A180005), Henan Province Science and Technology Project (grant number 222102310383), Doctoral Scientific Research Program Foundation of Xinxiang Medical University (grant number XYBSKYZZ201523), Research Innovation Support Program for Postgraduates of Xinxiang Medical University (grant number YJSCX202135Y).
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by Minlin Yang and Yanchun Zhao. The first draft of the manuscript was written by Minlin Yang and Lihong Guan, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
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Yang, M., Zhao, Y., Li, X. et al. Conditioned medium of human menstrual blood-derived endometrial stem cells protects against cell inflammation and apoptosis of Npc1KO N2a cells. Metab Brain Dis 38, 2301–2313 (2023). https://doi.org/10.1007/s11011-023-01243-1
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DOI: https://doi.org/10.1007/s11011-023-01243-1