Generation of hepatoma cell lines deficient in microsomal triglyceride transfer protein

Anaganti, Narasimha; Chattopadhyay, Atrayee; Poirier, John T.; Hussain, M. Mahmood

doi:10.1016/j.jlr.2022.100257

Cited by 7 publications

(4 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To understand the effect of I128T mutation on MTP activity, we generated plasmids expressing this mutation (Figures S4–S6) and expressed the mutant MTP or wild‐type (WT) MTP in MTTP knockout human hepatoma Huh‐7 (Mko‐3) cells 20 . The purified WT and I128T MTP proteins interacted with endogenous PDI to similar extents (Figure S5A).…”

Section: Resultsmentioning

confidence: 99%

“…Furthermore, I128T MTP was as potent as WT MTP protein in supporting the secretion of apoB100 (Figure S4B). We previously showed that MTP knockout cells accumulate lipids 20 . We tested if the expression of I128T protein would affect cellular lipid accumulation by transfecting Mko‐3 cells with either control, WT hMTP, or hI128T expression plasmids.…”

Section: Resultsmentioning

confidence: 99%

“…Site‐directed mutagenesis was used to generate the I128T mutation for expression of human MTP (hMTP) in MTP‐deficient human hepatoma Huh‐7 cells (‘Mko‐3’, Figures S4 and S5). 20 The ATC codon coding for Isoleucine at position 128 was targeted for a point mutation to ACC, which translates to Threonine. The wild‐type (WT) and I128T sequences were cloned into an expression vector with FLAG tags, and expression plasmids were sequenced to confirm the mutation (Figure S3).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A coding variant in the microsomal triglyceride transfer protein reduces both hepatic steatosis and plasma lipids

Schneider,

Hehl,

Creasy

et al. 2023

Aliment Pharmacol Ther

Self Cite

View full text Add to dashboard Cite

Summary Background Genetic inactivation and pharmacologic inhibition of the microsomal triglyceride transfer protein (MTP; gene name MTTP) inhibits hepatic secretion of VLDL, thereby reducing serum lipids and apoB at the expense of increasing hepatic steatosis. Aim To examine the effects of missense variants in MTTP on hepatic and circulating lipids. Methods We analysed the association of MTTP missense variants with metabolic, hepatic and clinical phenotypes in the Penn Medicine Biobank (PMBB; n = 37,960) and the UKBiobank (UKB; n = 451,444). Results We analysed 24 missense variants in MTTP in PMBB for association with biopsy‐proven hepatic steatosis and found that an isoleucine 128 to threonine variant (I128T: rs3816873‐A, frequency 26%) was associated with reduced steatosis (p < 0.001). PMBB subjects with imaging‐proven steatosis also revealed significantly fewer carriers of MTTP I128T compared to controls. Analysis in UKB also showed that MTTP I128T was associated with reduced risk of hepatic steatosis. Unexpectedly, MTTP I128T was found to be associated with reduced plasma levels of LDL‐cholesterol and apoB (all p < 0.001). Functional studies indicated that MTTP I128T is neither a classic loss nor gain of function allele. Conclusions MTTP I128T is associated with reduced hepatic steatosis as well as reduced plasma lipids and apoB. This paradoxical profile is not consistent with a simple gain or loss of function in MTP activity and suggests a more complex effect on MTP function. Further investigation of MTTP I128T will provide insight into the structure–function of MTP and potentially new approaches to modulate MTP activity that could both reduce hepatic and circulating lipids.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

A coding variant in the microsomal triglyceride transfer protein reduces both hepatic steatosis and plasma lipids

Schneider,

Hehl,

Creasy

et al. 2023

Aliment Pharmacol Ther

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human hepatoma Huh-7, mouse hepatocyte AML12, and mouse macrophage J774 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), and 1% L-glutamine in T75 flasks at 37°C in humidified 5% CO 2 incubators 25,26 .…”

Section: Methodsmentioning

confidence: 99%

MicroRNA-541-3p alters lipoproteins to reduce atherosclerosis by degrading Znf101 and Casz1 transcription factors

Ansari,

Yadav,

Zhou

et al. 2023

Preprint

View full text Add to dashboard Cite

High apoB-containing low-density lipoproteins (LDL) and low apoA1-containing high-density lipoproteins (HDL) are associated with atherosclerosis. In search of a molecular regulator that could simultaneously and reciprocally control both LDL and HDL levels, we screened a microRNA (miR) library using human hepatoma Huh-7 cells. We identified miR-541-3p that both decreases apoB and increases apoA1 expression by inducing mRNA degradation of two different transcription factors, Znf101 and Casz1. Znf101 enhances apoB expression while Casz1 represses apoA1 expression. The hepatic knockdown of orthologous Zfp961 and Casz1 genes in mice altered plasma lipoproteins and reduced atherosclerosis without causing hepatic lipid accumulation, most likely by lowering hepatic triglyceride production, increasing HDL cholesterol efflux capacity, and reducing lipogenesis. Notably, human genetic variants in the MIR541, ZNF101, and CASZ1 loci are significantly associated with plasma lipids and lipoprotein levels. This study identifies miR-541-3p and Znf101/Casz1 as potential therapeutic agent and targets, respectively, to reduce plasma lipoproteins and atherosclerosis without causing liver steatosis.

show abstract

New CRISPR Technology for Creating Cell Models of Lipoprotein Assembly and Secretion

Anaganti

Chattopadhyay

Filippo

et al. 2023

Curr Atheroscler Rep

View full text Add to dashboard Cite

Generation of hepatoma cell lines deficient in microsomal triglyceride transfer protein

Cited by 7 publications

References 37 publications

A coding variant in the microsomal triglyceride transfer protein reduces both hepatic steatosis and plasma lipids

A coding variant in the microsomal triglyceride transfer protein reduces both hepatic steatosis and plasma lipids

MicroRNA-541-3p alters lipoproteins to reduce atherosclerosis by degrading Znf101 and Casz1 transcription factors

New CRISPR Technology for Creating Cell Models of Lipoprotein Assembly and Secretion

Contact Info

Product

Resources

About