Home A single domain antibody against the Cys- and His-rich domain of PCSK9 and evolocumab exhibit different inhibition mechanisms in humanized PCSK9 mice
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A single domain antibody against the Cys- and His-rich domain of PCSK9 and evolocumab exhibit different inhibition mechanisms in humanized PCSK9 mice

  • Rachid Essalmani , Elodie Weider , Jadwiga Marcinkiewicz , Ann Chamberland , Delia Susan-Resiga , Anna Roubtsova , Nabil G. Seidah and Annik Prat EMAIL logo
Published/Copyright: August 31, 2018
Biological Chemistry
From the journal Biological Chemistry Volume 399 Issue 12

Abstract

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein that binds and escorts the low density lipoprotein receptor (LDLR) into the lysosomal degradation pathway. Prescribed monoclonal antibodies (mAbs) against PCSK9 prevent its binding to the LDLR, and result in ~60% lower LDL cholesterol (LDLc) levels. Although efficient, mAbs are expensive. Hence other PCSK9 inhibitors are needed. For screening purpose, we developed C57BL/6J mice expressing the human PCSK9 gene under the control of its own promoter, but lacking endogenous mouse PCSK9. All lines recapitulate the endogenous PCSK9 expression pattern. The Tg2 line that expresses physiological levels of human PCSK9 (hPCSK9) was selected to characterize the inhibitory properties of a previously reported single domain antibody (sdAb), PKF8-mFc, which binds the C-terminal domain of PCSK9. Upon intraveinous injection of 10 mg/kg, PKF8-mFc and the mAb evolocumab neutralized ~50% and 100% of the hPCSK9 impact on total cholesterol (TC) levels, respectively, but PKF8-mFc had a more sustained effect. PKF8-mFc barely affected hPCSK9 levels, whereas evolocumab promoted a 4-fold increase 3 days post-injection, suggesting very different inhibitory mechanisms. The present study also shows that the new transgenic mice are well suited to screen a variety of hPCSK9 inhibitors.

Keywords: cholesterol; familial hypercholesterolemia (FH); low-density lipoprotein (LDL); LDL receptor (LDLR); transgenic mouse

Acknowledgments

We would like to thank Manon Laprise for animal experimentation, Suzie Riverin for animal care, and Odile Neyret, Myriam Rondeau and Agnès Dumont for mouse genotyping and BAC copy numbers. This work was supported by the Canadian Institutes of Health Research grants Foundation Scheme, Funder Id: 10.13039/501100000035, 148363 and Funder Id: 10.13039/501100000029, MOP 102741, a Pfizer Aspire 1 cardiovascular research award, Funder Id: 10.13039/100004319, WI207162, a Canada Research Chair, Funder Id: 10.13039/501100001804, 231335, and a Fondation Leducq grant, Funder Id: 10.13039/501100001674, #13CVD03.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/hsz-2018-0194).

Received: 2018-03-20
Accepted: 2018-07-09
Published Online: 2018-08-31
Published in Print: 2018-11-27

©2018 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Highlight: Frontiers in Proteolysis
  3. Host cell-surface proteins as substrates of gingipains, the main proteases of Porphyromonas gingivalis
  4. A single domain antibody against the Cys- and His-rich domain of PCSK9 and evolocumab exhibit different inhibition mechanisms in humanized PCSK9 mice
  5. Characterization of PdCP1, a serine carboxypeptidase from Pseudogymnoascus destructans, the causal agent of White-nose Syndrome
  6. An internally quenched peptide as a new model substrate for rhomboid intramembrane proteases
  7. An alternative processing pathway of APP reveals two distinct cleavage modes for rhomboid protease RHBDL4
  8. Reviews
  9. Salivary peptide histatin 1 mediated cell adhesion: a possible role in mesenchymal-epithelial transition and in pathologies
  10. Modulation of dynamin function by small molecules
  11. Chemotherapeutic resistance: a nano-mechanical point of view
  12. Research Articles/Short Communications
  13. Protein Structure and Function
  14. Biochemical and kinetic properties of the complex Roco G-protein cycle
  15. Cell Biology and Signaling
  16. Aberrant expression of hsa_circ_0025036 in lung adenocarcinoma and its potential roles in regulating cell proliferation and apoptosis
https://doi.org/10.1515/hsz-2018-0194
Keywords for this article
cholesterol; familial hypercholesterolemia (FH); low-density lipoprotein (LDL); LDL receptor (LDLR); transgenic mouse

Articles in the same Issue

  1. Frontmatter
  2. Highlight: Frontiers in Proteolysis
  3. Host cell-surface proteins as substrates of gingipains, the main proteases of Porphyromonas gingivalis
  4. A single domain antibody against the Cys- and His-rich domain of PCSK9 and evolocumab exhibit different inhibition mechanisms in humanized PCSK9 mice
  5. Characterization of PdCP1, a serine carboxypeptidase from Pseudogymnoascus destructans, the causal agent of White-nose Syndrome
  6. An internally quenched peptide as a new model substrate for rhomboid intramembrane proteases
  7. An alternative processing pathway of APP reveals two distinct cleavage modes for rhomboid protease RHBDL4
  8. Reviews
  9. Salivary peptide histatin 1 mediated cell adhesion: a possible role in mesenchymal-epithelial transition and in pathologies
  10. Modulation of dynamin function by small molecules
  11. Chemotherapeutic resistance: a nano-mechanical point of view
  12. Research Articles/Short Communications
  13. Protein Structure and Function
  14. Biochemical and kinetic properties of the complex Roco G-protein cycle
  15. Cell Biology and Signaling
  16. Aberrant expression of hsa_circ_0025036 in lung adenocarcinoma and its potential roles in regulating cell proliferation and apoptosis
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