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Proteomics analysis of physiological effect of ZBTB16 (PLZF) on osteogenic differentiation of human valve interstitial cells (VICs) by DDA PASEF on TimsToF Pro

ABSTRACT: During fundamental proteotranscriptomic analysis of human valve interstitial cells (VICs) osteogenic differentiation, we revealed ZBTB16 (PLZF) as one of the most upregulated proteins. Thus, to investigate its physiological role in VICs osteogenic differentiation we performed its overexpression by lentiviral transduction with subsequent proteomics analysis. Despite obvious PLZF overexpression and its reinforcing effect on osteogenic differentiation, we revealed surprisingly small amount of differentially expressed proteins that assumed to be the targets of PLZF.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Heart Valve Cell

DISEASE(S): Aortic Valve Disease

SUBMITTER: Arseniy Lobov

LAB HEAD: Anna B. Malashicheva

PROVIDER: PXD031572 | Pride | 2022-10-21

REPOSITORIES: pride

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Project description:Mutations in the RMRP gene are the origin of cartilage-hair hypoplasia. Cartilage-hair hypoplasia is associated with severe dwarfism caused by impaired skeletal development. However, it is not clear why mutations in the RMRP gene lead to skeletal dysplasia. Viperin is a known substrate of RMRP. Since chondrogenic differentiation of the growth plate is required for development of the long bones, we hypothesized that viperin functions as a chondrogenic regulator downstream of RMRP. Viperin protein is expressed throughout the stages of chondrogenic differentiation in vivo. Viperin gene expression is increased during knockdown of Rmrp RNA in the ATDC5 model for chondrogenic differentiation. Viperin is expressed during ATDC5 chondrogenic differentiation. Viperin knockdown reduces, while viperin overexpression increases overall protein secretion, with CXCL10 identified as a potential target via mass spectrometry-proteomics. CXCL10 protein expression is reduced during knockdown and increased during overexpression of viperin and CXCL10 protein expression coincides with viperin expression in ATDC5 chondrogenic differentiation. Viperin knockdown induces, while viperin overexpression reduces TGFβ activity. Furthermore, viperin knockdown conditioned media increases, while viperin overexpression conditioned media reduces chondrogenic differentiation of ATDC5 cells. TGFβ target genes Pai1 and Smad7 are increased during knockdown and reduced during overexpression of viperin. Moreover, TGFβ activity is reduced when differentiating ATDC5 cells are exposed to CXCL10 and, acting as a viperin overexpression mimic, CXCL10 similarly reduces chondrogenic differentiation of ATDC5. Lastly, we show that in CHH patient cells, RMRP expression is reduced and viperin expression is increased, coinciding with reduced chondrogenic differentiation and increased CXCL10 expression, possibly explaining the CHH phenotype. Together our data show that viperin may play a pivotal role in chondrogenic differentiation, with potential consequences for cartilage-hair hypoplasia pathobiology.

			Action	DRS
	Contr_383.d.zip	Other
	Contr_383.mgf	Mgf
	Contr_482.d.zip	Other
	Contr_482.mgf	Mgf
	Contr_494.d.zip	Other

Dataset Information

Proteomics analysis of physiological effect of ZBTB16 (PLZF) on osteogenic differentiation of human valve interstitial cells (VICs) by DDA PASEF on TimsToF Pro

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