Mechanistic Toxicology Branch, Division of Translational Toxicology, National Institute of Environmental Health Sciences, Research Triangle Park, Durham, NC 27709, USA.
Molecular Genomics Core, Division of Intramural Research, National Institute of Environmental Health Sciences, Research Triangle Park, Durham, NC 27709, USA.
Int J Mol Sci. 2024 May 18;25(10):5522. doi: 10.3390/ijms25105522.
Early detection of neurological conditions is critical for timely diagnosis and treatment. Identifying cellular-level changes is essential for implementing therapeutic interventions prior to symptomatic disease onset. However, monitoring brain tissue directly through biopsies is invasive and poses a high risk. Bodily fluids such as blood or cerebrospinal fluid contain information in many forms, including proteins and nucleic acids. In particular, cell-free DNA (cfDNA) has potential as a versatile neurological biomarker. Yet, our knowledge of cfDNA released by brain tissue and how cfDNA changes in response to deleterious events within the brain is incomplete. Mapping changes in cfDNA to specific cellular events is difficult in vivo, wherein many tissues contribute to circulating cfDNA. Organoids are tractable systems for examining specific changes consistently in a human background. However, few studies have investigated cfDNA released from organoids. Here, we examined cfDNA isolated from cerebral organoids. We found that cerebral organoids release quantities of cfDNA sufficient for downstream analysis with droplet-digital PCR and whole-genome sequencing. Further, gene ontology analysis of genes aligning with sequenced cfDNA fragments revealed associations with terms related to neurodevelopment and autism spectrum disorder. We conclude that cerebral organoids hold promise as tools for the discovery of cfDNA biomarkers related to neurodevelopmental and neurological disorders.
早期发现神经疾病对于及时诊断和治疗至关重要。识别细胞水平的变化对于在症状出现前实施治疗干预至关重要。然而，通过活检直接监测脑组织是侵入性的，并且风险很高。体液，如血液或脑脊液，包含多种形式的信息，包括蛋白质和核酸。特别是，无细胞 DNA（cfDNA）作为一种多功能神经生物标志物具有潜力。然而，我们对脑组织释放的 cfDNA 以及 cfDNA 如何响应大脑内有害事件而发生变化的了解还不完整。将 cfDNA 的变化映射到特定的细胞事件在体内是困难的，因为许多组织都有助于循环 cfDNA。类器官是在人类背景下一致检查特定变化的可行系统。然而，很少有研究调查从类器官释放的 cfDNA。在这里，我们检查了从大脑类器官中分离出的 cfDNA。我们发现大脑类器官释放的 cfDNA 数量足以进行后续的液滴数字 PCR 和全基因组测序分析。此外，与测序 cfDNA 片段对齐的基因的基因本体分析显示与神经发育和自闭症谱系障碍相关的术语相关联。我们得出结论，大脑类器官有望成为发现与神经发育和神经紊乱相关的 cfDNA 生物标志物的工具。
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