PXD063568

PXD063568 is an original dataset announced via ProteomeXchange.

Title	Proteomic Landscape of Porcine Induced Neural Stem Cell Reprogramming and Differentiation
Description	Background: Direct reprogramming of somatic cells into induced neural stem cells (iNSCs) holds strong potential for regenerative medicine, especially in large animal models like pigs, which are crucial for translational and preclinical research. However, the molecular mechanisms driving porcine fibroblast-to-iNSC conversion and subsequent differentiation remain poorly understood at the proteomic level. Methods: To map the proteomic changes during reprogramming and differentiation, we performed unbiased label-free discovery proteomics (nano-LC-MS/MS) and targeted SWATH-MS quantification. Proteomes of porcine tail fibroblasts (PTFs), porcine iNSCs (piNSCs; passage 20), and their neuronal and glial differentiated progeny (piNSCs-NGs; passage 21, after 14 days of neural induction) were compared. Two previously established piNSC lines (VSMUi002-B and VSMUi002-E), generated via Sendai virus-mediated reprogramming, were used as the cellular models. Results: The piNSC lines displayed hallmark NSC morphology and expressed canonical markers (PAX6+, SOX2+, NES+, VIM+, OCT4−). Upon differentiation, they generated neuronal and glial cells expressing TUJ1, MAP2, SYP, TH, and GFAP, confirming their multipotency. A total of 4,094 proteins were identified across the three cell states. Multivariate analysis revealed distinct proteomic signatures separating fibroblasts, iNSCs, and their neuronal/glial progeny. The transition from fibroblast to piNSC was marked by increased expression of STMN1, LIMA1, TKT, and NEFL alongside suppression of ARPC5, ACO2, ETFB, and ALDOB. These shifts indicate profound cytoskeletal remodeling and metabolic reprogramming, reflecting a loss of fibroblast identity and the acquisition of a NSC state. During piNSC-to-piNSCs-NGs differentiation, 19 proteins were consistently upregulated. These included neuronal structural proteins (INA, STMN1), cytoskeletal regulators (PFN1), signaling modulators (MBIP), and proteins involved in lysosomal function (NCOA7), cell adhesion (CDHR2), and calcium signaling (ANXA4). Pathway and network analyses highlighted post-transcriptional regulation—particularly involving RNA processing and the RNA exosome complex (e.g., EXOSC3)—as a key feature of differentiation. Conclusion: This study provides the first comprehensive proteomic map of piNSC reprogramming and differentiation in a large animal model. Our findings uncover critical regulatory proteins and pathways governing cytoskeletal organization, metabolism, and RNA processing, offering valuable insights into neural fate transitions. This resource advances the understanding of neural reprogramming in translational models and supports future regenerative and comparative neuroscience efforts.
HostingRepository	PRIDE
AnnounceDate	2025-05-26
AnnouncementXML	Submission_2025-05-26_07:18:34.742.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sataporn Phochantachinda
SpeciesList	scientific name: Sus scrofa domesticus (domestic pig); NCBI TaxID: 9825;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	compact

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-02 12:26:19	ID requested
⏵ 1	2025-05-26 07:18:35	announced

Dataset with its publication pending

submitter keyword: piNSCs

porcine

proteomics

direct reprogramming

neuronal differentiation

mass spectrometry,induced neural stem cells

Piyada Na
contact affiliation	Center for Research Innovation and Bioinformatics, Faculty of Medical Technology, Mahidol University, Nakhon Pathom, Thailand
contact email	Piyada.naa@mahidol.ac.th
lab head
Sataporn Phochantachinda
contact affiliation	Faculty of Veterinary Science, Mahidol University
contact email	sataporn.pho@mahidol.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD063568
     1. Label: PRIDE project
     2. Name: Proteomic Landscape of Porcine Induced Neural Stem Cell Reprogramming and Differentiation