Lye Jed J, Williams Anthony, Baralle Diana
University of Southampton Medical School, University of Southampton, Southampton, United Kingdom.
Wessex Investigational Sciences Hub Laboratory (WISH Lab), Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
Front Genet. 2019 Dec 11;10:1204. doi: 10.3389/fgene.2019.01204. eCollection 2019.
Challenges in diagnosing primary immunodeficiency are numerous and diverse, with current whole-exome and whole-genome sequencing approaches only able to reach a molecular diagnosis in 25-60% of cases. We assess these problems and discuss how RNA-focused analysis has expanded and improved in recent years and may now be utilized to gain an unparalleled insight into cellular immunology. We review how investigation into RNA biology can give information regarding the differential expression, monoallelic expression, and alternative splicing-which have important roles in immune regulation and function. We show how this information can inform bioinformatic analysis pipelines and aid in the variant filtering process, expediting the identification of causal variants-especially those affecting splicing-and enhance overall diagnostic ability. We also demonstrate the challenges, which remain in the design of this type of investigation, regarding technological limitation and biological considerations and suggest potential directions for the clinical applications.
原发性免疫缺陷的诊断面临诸多挑战，且形式多样，目前的全外显子组测序和全基因组测序方法仅能在25%至60%的病例中实现分子诊断。我们评估了这些问题，并讨论了近年来以RNA为重点的分析是如何扩展和改进的，以及现在如何利用它来获得对细胞免疫学的无与伦比的见解。我们回顾了对RNA生物学的研究如何能够提供有关差异表达、单等位基因表达和可变剪接的信息，这些在免疫调节和功能中具有重要作用。我们展示了这些信息如何为生物信息学分析流程提供参考，并有助于变异筛选过程，加速因果变异的识别，尤其是那些影响剪接的变异，并提高整体诊断能力。我们还展示了这类研究设计中在技术限制和生物学考量方面仍然存在的挑战，并提出了临床应用的潜在方向。
