Dataset Information

Proteomic and phosphoproteomic profiling of SFTPCI73T expressing patient-specific pluripotent stem cell-derived alveolar epithelial type 2 cells and their gene-edited corrected counterparts.

ABSTRACT: We performed proteomic and phosphoproteomic profiling of cells derived from human induced pluripotent stem cells (iPSCs) using our previously described distal lung directed differentiation protocol to generate alveolar epithelial type 2 cells (iAEC2s). We used the SPC2 human iPSC line and specifically the SPC2-ST-B2 (SFTPCtdT/WT) and SPC2-ST-C11 (SFTPCI73T/tdT) clones containing a SFTPCtdTomato knock-in reporter. SFTPCtdTomato+ cells were sorted on day 41 and again on day 79 of differentiation. iAEC2s were single-cell passaged in self-renewing 3D alveolosphere cultures approximately every 2 weeks through day 113. Live SFTPCtdTomato+ iAEC2s were sorted on day 113 and processed for mass spectrometry. We find that mutant (SFTPCI73T/tdT) iAEC2s display a less proliferative and more mature AEC2 phenotype compared to their corrected (SFTPCtdT/WT) counterparts with a concomitant upregulation of lysosomal and autophagy related pathways and activation of the NF-κB pathway in mutant cells.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Type Ii Pneumocyte

DISEASE(S): Interstitial Lung Disease

SUBMITTER: Ryan Hekman

LAB HEAD: Darrell N. Kotton

PROVIDER: PXD022334 | Pride | 2021-10-07

REPOSITORIES: pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	20200216_RH_KR_F10_Phos.raw	Raw
	20200216_RH_KR_F10_Prot.raw	Raw
	20200216_RH_KR_F11_Phos.raw	Raw
	20200216_RH_KR_F11_Prot.raw	Raw
	20200216_RH_KR_F12_Phos.raw	Raw

Items per page:

1 - 5 of 34

Similar Datasets

Project description:We show that high quality microarray gene expression profiles can be obtained following FACS sorting of cells using combinations of transcription factors. We use this transcription factor FACS (tfFACS) methodology to perform a genomic analysis of hESC-derived endodermal lineages marked by combinations of SOX17, GATA4, and CXCR4, and find that triple positive cells have a much stronger definitive endoderm signature than other combinations of these markers. Additionally, SOX17+GATA4+ cells can be obtained at a much earlier stage of differentiation, prior to expression of CXCR4+ cells, providing an important new tool to isolate this earlier definitive endoderm subtype. Overall, tfFACS represents an advancement in FACS technology which broadly crosses multiple disciplines, most notably in regenerative medicine to redefine cellular populations. 21 Total samples were analyzed. Samples collected after 5 days of hESC differentiation included SOX17+GATA4+ CXCR4+ cells, unfixed CXCR4+ cells, and unsorted fixed cells. Samples collected after 3 days of differentiation included SOX17+GATA4+ cells, SOX17-GATA- cells, and unsorted fixed cells. As controls, we analyzed both unfixed hESCs and fixed hESCs. All samples contained biological duplicates, triplicates or quadriplicates. We performed hierarchical clustering to demonstrate whether cellular fixation alone could change gene expression. We based this analysis on 1647 transcript clusters with coefficient of variation > 0.5 across the samples and expression values >= 500 in at least 2 out of the 21 samples. We found that fixation and staining steps did not cause distortions in gene expression measurements. This is supported by the fact that fixed and unfixed cells cluster together based upon differentiation stage, not based upon degree of fixation. Furthermore, using GSEA analysis, we found that the SOX17+GATA4+CXCR4+ day 5 cells and day 3 SOX17+GATA4+ were more enriched for compiled gold-standard endodermal genes than the CXCR4+ day 5 population.

Dataset Information

Proteomic and phosphoproteomic profiling of SFTPCI73T expressing patient-specific pluripotent stem cell-derived alveolar epithelial type 2 cells and their gene-edited corrected counterparts.

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets