Single-Cell Transcriptional Profiling of Cells Derived From Regenerating Alveolar Ducts

Frontiers in Medicine

article

Alexandra B. Ysasi¹  Prapti Pokharel²  Kenneth J. Livak¹  Maximilian Ackermann³  Paul C. Blainey²  Steven J. Mentzer¹  Robert D. Bennett¹  Willi Wagner³  Cristian D. Valenzuela¹  Andrew B. Servais¹  Akira Tsuda⁵  Saumyadipta Pyne⁶  Shuqiang Li¹  Jonna Grimsby² 

[1] Laboratory of Adaptive and Regenerative Biology, Harvard Medical School, Brigham & Women's Hospital, United States;Broad Institute of Harvard and MIT, United States;Institute of Functional and Clinical Anatomy, University Medical Center of the Johannes Gutenberg-University;Department of Biological Engineering, United States;Molecular and Integrative Physiological Sciences, Harvard School of Public Health, United States;Public Health Dynamics Laboratory, University of Pittsburgh, United States

关键词: warburg effect;    glucose metabolism;    aerobic glycolysis;    metabolic reprogramming;    cholangiocarcinoma;   

DOI  :  10.3389/fmed.2020.00112

学科分类：社会科学、人文和艺术（综合）

来源: Frontiers

【 摘 要 】

Lung regeneration occurs in a variety of adult mammals after surgical removal of one lung (pneumonectomy). Previous studies of murine post-pneumonectomy lung growth have identified regenerative “hotspots” in subpleural alveolar ducts; however, the cell-types participating in this process remain unclear. To identify the single cells participating in post-pneumonectomy lung growth, we used laser microdissection, enzymatic digestion and microfluidic isolation. Single-cell transcriptional analysis of the murine alveolar duct cells was performed using the C1 integrated fluidic circuit (Fluidigm) and a custom PCR panel designed for lung growth and repair genes. The multi-dimensional data set was analyzed using visualization software based on the tSNE algorithm. The analysis identified 6 cell clusters; 1 cell cluster was present only after pneumonectomy. This post-pneumonectomy cluster was significantly less transcriptionally active than 3 other clusters and may represent a transitional cell population. A provisional cluster identity for 4 of the 6 cell clusters was obtained by embedding bulk transcriptional data into the tSNE analysis. The transcriptional pattern of the 6 clusters was further analyzed for genes associated with lung repair, matrix production, and angiogenesis. The data demonstrated that multiple cell-types (clusters) transcribed genes linked to these basic functions. We conclude that the coordinated gene expression across multiple cell clusters is likely a response to a shared regenerative microenvironment within the subpleural alveolar ducts.

【 授权许可】

CC BY