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SiRNA-mediated knockdown of StARD phospholipid transfer proteins in HepG2 cells

ABSTRACT: We used siRNA to individually knock down the human phospholipid transfer proteins STARD2, STARD7 and STARD10 in HepG2 cells and measured the impact on the cellular phospholipid metabolism. We used DIA proteomics to quantify the selective reduction in protein expression levels.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hepatocyte, Cell Culture

SUBMITTER: Carla Kirschbaum

LAB HEAD: Carol Robinson

PROVIDER: PXD072869 | Pride | 2026-07-21

REPOSITORIES: Pride

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			Action	DRS
	2025-10-30-decoys-reviewed-contam-UP000005640.fas	Other
	OR_10_1_Slot2-34_6211.d.zip	Other
	OR_10_2_Slot2-35_6212.d.zip	Other
	OR_10_3_Slot2-36_6213.d.zip	Other
	OR_10_4_Slot2-43_6214.d.zip	Other

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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.

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SiRNA-mediated knockdown of StARD phospholipid transfer proteins in HepG2 cells

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