Identifying and characterizing lung progenitor cells and their signaling niches is crucial for understanding how a healthy lung is built and maintained, how alteration of development and maintenance pathways cause or contribute to lung disease, and how disease can be prevented and damaged lung tissue restored or replaced. Identifying and characterizing lung progenitor cells and their niches has been hampered by the complex three-dimensional structure of the lung and the lack of tools to mark, follow the fate, and manipulate gene expression in individual lung cells in vivo. Here we propose to develop such tools, by adapting for use in mouse lung the systematic genetic approaches and single cell resolution genetic tools ("clonal analysis") that have been used over the past decade to elucidate progenitor and stem cells and their signaling niches in the model organism Drosophila. To facilitate this, we will develop a new microscopy procedure (multidimensional microscopic molecular profiling) that allows the levels of dozens of molecular markers to be quantified at subcellular resolution on a single tissue section, the equivalent of multi-dimensional FACS analysis for intact tissues. We combine these in vivo approaches with a high throughput in vitro approach that takes advantage of recent advances in genomics and microfluidics to characterize individual lung progenitor cells and their developmental responses to the signals identified in the in vivo experiments. This combined approach is general and applicable to progenitor cells throughout the lung and other mouse tissues, although we focus on the poorly characterized progenitor cells in lung mesenchyme.

Agency
National Institute of Health (NIH)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HL099995-06
Application #
8656740
Study Section
Special Emphasis Panel (ZHL1)
Program Officer
Blaisdell, Carol J
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Stanford University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Stanford
State
CA
Country
United States
Zip Code
94304
Alanis, Denise Martinez; Chang, Daniel R; Akiyama, Haruhiko et al. (2014) Two nested developmental waves demarcate a compartment boundary in the mouse lung. Nat Commun 5:3923
Desai, Tushar J; Brownfield, Douglas G; Krasnow, Mark A (2014) Alveolar progenitor and stem cells in lung development, renewal and cancer. Nature 507:190-4
Wang, Jianbin; Quake, Stephen R (2014) RNA-guided endonuclease provides a therapeutic strategy to cure latent herpesviridae infection. Proc Natl Acad Sci U S A 111:13157-62
Wu, Angela R; Neff, Norma F; Kalisky, Tomer et al. (2014) Quantitative assessment of single-cell RNA-sequencing methods. Nat Methods 11:41-6
Treutlein, Barbara; Brownfield, Doug G; Wu, Angela R et al. (2014) Reconstructing lineage hierarchies of the distal lung epithelium using single-cell RNA-seq. Nature 509:371-5
Riegler, Johannes; Gillich, Astrid; Shen, Qi et al. (2014) Cardiac tissue slice transplantation as a model to assess tissue-engineered graft thickness, survival, and function. Circulation 130:S77-86
Kumar, Maya E; Bogard, Patrick E; Espinoza, F HernĂ¡n et al. (2014) Mesenchymal cells. Defining a mesenchymal progenitor niche at single-cell resolution. Science 346:1258810
Tan, Fraser E; Vladar, Eszter K; Ma, Lina et al. (2013) Myb promotes centriole amplification and later steps of the multiciliogenesis program. Development 140:4277-86