RNA therapeutics inactivate PCSK9 by inducing a unique intracellular retention form

Rocha, Cristina S. J.; Wiklander, Oscar P. B.; Larsson, Lars-Olof; Moreno, Pedro M. D.; Parini, Paolo; Lundin, Karin E.; Smith, C. I. Edvard

doi:10.1016/j.yjmcc.2015.03.009

Cited by 19 publications

(16 citation statements)

References 47 publications

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“…S3E). Increased ER expansion, another hallmark feature of ER stress, was also observed in HepG2 cells transfected with a splice-switching oligomer that causes ER retention of PCSK9 in response to PA 25 (Fig. S4A and S4B).…”

Section: Pcsk9 Protects Cultured Hepatocytes From Palmitate-induced Ementioning

confidence: 63%

Pcsk9 knockout exacerbates diet-induced non-alcoholic steatohepatitis, fibrosis and liver injury in mice

et al. 2019

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Background & Aims: The fatty acid translocase, also known as CD36, is a well-established scavenger receptor for fatty acid (FA) uptake and is abundantly expressed in many metabolically active tissues. In the liver, CD36 is known to contribute to the progression of non-alcoholic fatty liver disease and to the more severe non-alcoholic steatohepatitis, by promoting triglyceride accumulation and subsequent lipid-induced endoplasmic reticulum (ER) stress. Given the recent discovery that the hepatocyte-secreted proprotein convertase subtilisin/kexin type 9 (PCSK9) blocks CD36 expression, we sought to investigate the role of PCSK9 in liver fat accumulation and injury in response to saturated FAs and in a mouse model of diet-induced hepatic steatosis. Methods: In this study, we investigated the role of PCSK9 on the uptake and accumulation of FAs, as well as FA-induced toxicity, in a variety of cultured hepatocytes. Diet-induced hepatic steatosis and liver injury were also assessed in Pcsk9-/mice. Results: Our results indicate that PCSK9 deficiency in cultured hepatocytes increased the uptake and accumulation of saturated and unsaturated FAs. In the presence of saturated FAs, PCSK9 also protected cultured hepatocytes from ER stress and cytotoxicity. In line with these findings, a metabolic challenge using a high-fat diet caused severe hepatic steatosis, ER stress inflammation and fibrosis in the livers of Pcsk9-/mice compared to controls. Given that inhibition of CD36 ablated the observed accumulation of lipid in vitro and in vivo, our findings also highlight CD36 as a strong contributor to steatosis and liver injury in the context of PCSK9 deficiency. Conclusions: Collectively, our findings demonstrate that PCSK9 regulates hepatic triglyceride content in a manner dependent on CD36. In the presence of excess dietary fats, PCSK9 can also protect against hepatic steatosis and liver injury.

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Section: Pcsk9 Protects Cultured Hepatocytes From Palmitate-induced Ementioning

confidence: 63%

Pcsk9 knockout exacerbates diet-induced non-alcoholic steatohepatitis, fibrosis and liver injury in mice

et al. 2019

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“…Aside from antibodies that have received approval (38), there is considerable interest in identifying other types of PCSK9 antagonists as potential therapeutics (42–44). To identify a SOMAmer inhibitor of PCSK9, 41 SOMAmers (30-mers, K d < 1 nM) were screened in a plate-based sandwich assay where biotinylated PCSK9 was incubated with or without SOMAmer, added to an LDL-R–coated plate, and detected using streptavidin-HRP conjugate in a chemiluminescent readout ( SI Appendix , Fig.…”

Section: Resultsmentioning

confidence: 99%

Selection of DNA aptamers with two modified bases

Gawande

Rohloff

Carter

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

169

139

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SignificanceAptamers are now used ubiquitously as binding agents for a broad range of applications. Natural (unmodified) DNA and RNA aptamers have considerably less chemical diversity than protein-based ligands such as antibodies, limiting their utility. Aptamers possessing a single chemical modification have helped bridge this diversity gap. We report the selection and identification of aptamers with two diversity-enhancing chemical modifications that bind and inhibit proprotein convertase subtilisin/kexin type 9 (PCSK9), a representative human therapeutic protein target. The addition of a second modification, especially in certain pairwise combinations, resulted in significant improvements in affinity, ligand efficiency, epitope coverage, metabolic stability, and inhibitory activity. Extensively chemically functionalized aptamers have the potential to become the next generation of nucleic-acid–based ligands.

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“…Following administration of SSO, an increase of the selected splice form at both the mRNA and protein level was detected when compared to nontreated Huh7 and HepG2 cell lines, with a concomitant increase of the LDL-R protein level. In vivo, full conversion to the splice form was achieved in a reporter system when mice were treated with the specific SSO [156]. …”

Section: Approaches To Reduce Pcsk9 Synthesis And/or Pcsk9/ldlr Intermentioning

confidence: 99%

Physiological and therapeutic regulation of PCSK9 activity in cardiovascular disease

et al. 2017

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Ischemic heart disease is the main cause of death worldwide and is accelerated by increased levels of low-density lipoprotein cholesterol (LDL-C). Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a potent circulating regulator of LDL-C through its ability to induce degradation of the LDL receptor (LDLR) in the lysosome of hepatocytes. Only in the last few years, a number of breakthroughs in the understanding of PCSK9 biology have been reported illustrating how PCSK9 activity is tightly regulated at several levels by factors influencing its transcription, secretion, or by extracellular inactivation and clearance. Two humanized antibodies directed against the LDLR-binding site in PCSK9 received approval by the European and US authorities and additional PCSK9 directed therapeutics are climbing up the phases of clinical trials. The first outcome data of the PCSK9 inhibitor evolocumab reported a significant reduction in the composite endpoint (cardiovascular death, myocardial infarction, or stroke) and further outcome data are awaited. Meanwhile, it became evident that PCSK9 has (patho)physiological roles in several cardiovascular cells. In this review, we summarize and discuss the recent biological and clinical data on PCSK9, the regulation of PCSK9, its extra-hepatic activities focusing on cardiovascular cells, molecular concepts to target PCSK9, and finally briefly summarize the data of recent clinical studies.

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RNA therapeutics inactivate PCSK9 by inducing a unique intracellular retention form

Cited by 19 publications

References 47 publications

Pcsk9 knockout exacerbates diet-induced non-alcoholic steatohepatitis, fibrosis and liver injury in mice

Pcsk9 knockout exacerbates diet-induced non-alcoholic steatohepatitis, fibrosis and liver injury in mice

Selection of DNA aptamers with two modified bases

Physiological and therapeutic regulation of PCSK9 activity in cardiovascular disease

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