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Novel Apo E-Derived ABCA1 Agonist Peptide (CS-6253) Promotes Reverse Cholesterol Transport and Induces Formation of preβ-1 HDL In Vitro.

Hafiane A, Bielicki JK, Johansson JO, Genest J - PLoS ONE (2015)

Bottom Line: We have previously reported the effects of compound ATI-5261 on its ability to replicate many functions of native apo A-I in the process of HDL biogenesis.CS-6253 significantly increases cholesterol efflux in murine macrophages and in human THP-1 macrophage-derived foam cells expressing ABCA1.When incubated with human plasma CS-6253 was also found to bind with HDL and LDL and promoted the transfer of cholesterol from HDL to LDL predominantly.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Laboratories Laboratory, Research Institute of the McGill University Health Centre, Montréal, Québec H4A 3J1, Canada.

ABSTRACT
Apolipoprotein (apo) mimetic peptides replicate some aspects of HDL function. We have previously reported the effects of compound ATI-5261 on its ability to replicate many functions of native apo A-I in the process of HDL biogenesis. ATI-5261 induced muscle toxicity in wild type C57Bl/6 mice, increased CPK, ALT and AST and increase in triglyceride (Tg) levels. Aromatic phenylalanine residues on the non-polar face of ATI-5261, together with positively charged arginine residues at the lipid-water interface were responsible for these effects. This information was used to create a novel analog (CS-6253) that was non-toxic. We evaluated this peptide designed from the carboxyl terminus of apo E, in its ability to mimic apo A-I functionality. Our data shows that the lipidated particles generated by incubating cells overexpressing ABCA1 with lipid free CS-6253 enhances the rate of ABCA1 lipid efflux with high affinity interactions with native ABCA1 oligomeric forms and plasma membrane micro-domains. Interaction between ABCA1 and lipid free CS-6253 resulted in formation of nascent HDL-CS-6253 particles that are actively remodeled in plasma. Mature HDL-CS-6253 particles deliver cholesterol to liver cells via SR-BI in-vitro. CS-6253 significantly increases cholesterol efflux in murine macrophages and in human THP-1 macrophage-derived foam cells expressing ABCA1. Addition of CS-6253 to plasma dose-dependently displaced apo A-I from α-HDL particles and led to de novo formation of preβ-1 HDL that stimulates ABCA1 dependent cholesterol efflux efficiently. When incubated with human plasma CS-6253 was also found to bind with HDL and LDL and promoted the transfer of cholesterol from HDL to LDL predominantly. Our data shows that CS-6253 mimics apo A-I in its ability to promote ABCA1-mediated formation of nascent HDL particles, and enhances formation of preβ-1 HDL with increase in the cycling of apo A-I between the preβ and α-HDL particles in-vitro. These mechanisms are potentially anti-atherogenic.

No MeSH data available.


Related in: MedlinePlus

Time course ability of CS-6253 to generate nHDL-mimetic like particles.(A and B): nHDL-CS-6253 and nHDL-apo A-I released to the medium at specific time points (45 min and 6 h) from BHK cells expressing ABCA1 were analyzed by 2D-PAGGE. Apo A-I was detected by polyclonal anti-apo A-I, CS-6253 peptides was detected by anti-CS-6253 and revealed by chemiluminescence. Molecular markers are indicated.
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pone.0131997.g005: Time course ability of CS-6253 to generate nHDL-mimetic like particles.(A and B): nHDL-CS-6253 and nHDL-apo A-I released to the medium at specific time points (45 min and 6 h) from BHK cells expressing ABCA1 were analyzed by 2D-PAGGE. Apo A-I was detected by polyclonal anti-apo A-I, CS-6253 peptides was detected by anti-CS-6253 and revealed by chemiluminescence. Molecular markers are indicated.

Mentions: Next, we investigated the nature of CS-6253-containing particles released in the medium from stimulated ABCA1 cells. Cells were first incubated in 100 mm diameter dishes with either (0.96 μM) apo A-I or CS-6253 in 8 ml of DMEM for 45 min and 6 h at 37°C. These two time points were selected based on our previous work on HDL biogenesis [25, 36]. Lipid free apo A-I or CS-6253 incubated in cell free system media under same conditions were used as control for 2D-PAGGE analysis. Apo A-I or CS-6253-containing particles were separated and analyzed by 2D-PAGGE. As shown in Fig 5, left panel, apo A-I or CS-6253 incubated without cells had a pre-β electrophoretic mobility with a molecular diameter of 7.1 nm. However, apo A-I and CS-6253-containing particles released from stimulated ABCA1 cells at either 45min or 6 h exhibited α-electrophoretic mobility with a particle size ranging from 9 to 20 nm (designated α-LpA-I-like particles, α-LpCS-6253-like particles) (Fig 5A and 5B) respectively. Both the charge and size of these nascent particles were stable over a 6h incubation period. These nHDL particles were retained following 50 kDa molecular weight cut filter separation, suggesting a particle size that is equal to, or greater than 20 nm nHDL-LpA-I [21].


Novel Apo E-Derived ABCA1 Agonist Peptide (CS-6253) Promotes Reverse Cholesterol Transport and Induces Formation of preβ-1 HDL In Vitro.

Hafiane A, Bielicki JK, Johansson JO, Genest J - PLoS ONE (2015)

Time course ability of CS-6253 to generate nHDL-mimetic like particles.(A and B): nHDL-CS-6253 and nHDL-apo A-I released to the medium at specific time points (45 min and 6 h) from BHK cells expressing ABCA1 were analyzed by 2D-PAGGE. Apo A-I was detected by polyclonal anti-apo A-I, CS-6253 peptides was detected by anti-CS-6253 and revealed by chemiluminescence. Molecular markers are indicated.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4514675&req=5

pone.0131997.g005: Time course ability of CS-6253 to generate nHDL-mimetic like particles.(A and B): nHDL-CS-6253 and nHDL-apo A-I released to the medium at specific time points (45 min and 6 h) from BHK cells expressing ABCA1 were analyzed by 2D-PAGGE. Apo A-I was detected by polyclonal anti-apo A-I, CS-6253 peptides was detected by anti-CS-6253 and revealed by chemiluminescence. Molecular markers are indicated.
Mentions: Next, we investigated the nature of CS-6253-containing particles released in the medium from stimulated ABCA1 cells. Cells were first incubated in 100 mm diameter dishes with either (0.96 μM) apo A-I or CS-6253 in 8 ml of DMEM for 45 min and 6 h at 37°C. These two time points were selected based on our previous work on HDL biogenesis [25, 36]. Lipid free apo A-I or CS-6253 incubated in cell free system media under same conditions were used as control for 2D-PAGGE analysis. Apo A-I or CS-6253-containing particles were separated and analyzed by 2D-PAGGE. As shown in Fig 5, left panel, apo A-I or CS-6253 incubated without cells had a pre-β electrophoretic mobility with a molecular diameter of 7.1 nm. However, apo A-I and CS-6253-containing particles released from stimulated ABCA1 cells at either 45min or 6 h exhibited α-electrophoretic mobility with a particle size ranging from 9 to 20 nm (designated α-LpA-I-like particles, α-LpCS-6253-like particles) (Fig 5A and 5B) respectively. Both the charge and size of these nascent particles were stable over a 6h incubation period. These nHDL particles were retained following 50 kDa molecular weight cut filter separation, suggesting a particle size that is equal to, or greater than 20 nm nHDL-LpA-I [21].

Bottom Line: We have previously reported the effects of compound ATI-5261 on its ability to replicate many functions of native apo A-I in the process of HDL biogenesis.CS-6253 significantly increases cholesterol efflux in murine macrophages and in human THP-1 macrophage-derived foam cells expressing ABCA1.When incubated with human plasma CS-6253 was also found to bind with HDL and LDL and promoted the transfer of cholesterol from HDL to LDL predominantly.

View Article: PubMed Central - PubMed

Affiliation: Cardiovascular Research Laboratories Laboratory, Research Institute of the McGill University Health Centre, Montréal, Québec H4A 3J1, Canada.

ABSTRACT
Apolipoprotein (apo) mimetic peptides replicate some aspects of HDL function. We have previously reported the effects of compound ATI-5261 on its ability to replicate many functions of native apo A-I in the process of HDL biogenesis. ATI-5261 induced muscle toxicity in wild type C57Bl/6 mice, increased CPK, ALT and AST and increase in triglyceride (Tg) levels. Aromatic phenylalanine residues on the non-polar face of ATI-5261, together with positively charged arginine residues at the lipid-water interface were responsible for these effects. This information was used to create a novel analog (CS-6253) that was non-toxic. We evaluated this peptide designed from the carboxyl terminus of apo E, in its ability to mimic apo A-I functionality. Our data shows that the lipidated particles generated by incubating cells overexpressing ABCA1 with lipid free CS-6253 enhances the rate of ABCA1 lipid efflux with high affinity interactions with native ABCA1 oligomeric forms and plasma membrane micro-domains. Interaction between ABCA1 and lipid free CS-6253 resulted in formation of nascent HDL-CS-6253 particles that are actively remodeled in plasma. Mature HDL-CS-6253 particles deliver cholesterol to liver cells via SR-BI in-vitro. CS-6253 significantly increases cholesterol efflux in murine macrophages and in human THP-1 macrophage-derived foam cells expressing ABCA1. Addition of CS-6253 to plasma dose-dependently displaced apo A-I from α-HDL particles and led to de novo formation of preβ-1 HDL that stimulates ABCA1 dependent cholesterol efflux efficiently. When incubated with human plasma CS-6253 was also found to bind with HDL and LDL and promoted the transfer of cholesterol from HDL to LDL predominantly. Our data shows that CS-6253 mimics apo A-I in its ability to promote ABCA1-mediated formation of nascent HDL particles, and enhances formation of preβ-1 HDL with increase in the cycling of apo A-I between the preβ and α-HDL particles in-vitro. These mechanisms are potentially anti-atherogenic.

No MeSH data available.


Related in: MedlinePlus