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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): Q Exactive HF-X

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

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			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

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

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