Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells
Delivering nucleic acids into the endothelium has great potential in treating vascular diseases. However, endothelial cells, which line the vasculature, are considered as sensitive in nature and hard to transfect. Low transfection efficacies in endothelial cells limit their potential therapeutic app...
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MDPI AG
2021-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/15/4626 |
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| author | Santhosh Chandar Maddila Chandrashekhar Voshavar Porkizhi Arjunan Rashmi Prakash Chowath Hari Krishna Reddy Rachamalla Balaji Balakrishnan Poonkuzhali Balasubramanian Rajkumar Banerjee Srujan Marepally |
| author_facet | Santhosh Chandar Maddila Chandrashekhar Voshavar Porkizhi Arjunan Rashmi Prakash Chowath Hari Krishna Reddy Rachamalla Balaji Balakrishnan Poonkuzhali Balasubramanian Rajkumar Banerjee Srujan Marepally |
| author_sort | Santhosh Chandar Maddila |
| collection | DOAJ |
| description | Delivering nucleic acids into the endothelium has great potential in treating vascular diseases. However, endothelial cells, which line the vasculature, are considered as sensitive in nature and hard to transfect. Low transfection efficacies in endothelial cells limit their potential therapeutic applications. Towards improving the transfection efficiency, we made an effort to understand the internalization of lipoplexes into the cells, which is the first and most critical step in nucleic acid transfections. In this study, we demonstrated that the transient modulation of caveolae/lipid rafts mediated endocytosis with the cholesterol-sequestrating agents, nystatin, filipin III, and siRNA against Cav-1, which significantly increased the transfection properties of cationic lipid-(2-hydroxy-<i>N</i>-methyl-<i>N</i>,<i>N</i>-bis(2-tetradecanamidoethyl)ethanaminium chloride), namely, amide liposomes in combination with 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) (AD Liposomes) in liver sinusoidal endothelial cells (SK-Hep1). In particular, nystatin was found to be highly effective with 2–3-fold enhanced transfection efficacy when compared with amide liposomes in combination with Cholesterol (AC), by switching lipoplex internalization predominantly through clathrin-mediated endocytosis and macropinocytosis. |
| first_indexed | 2024-03-10T09:11:04Z |
| format | Article |
| id | doaj.art-5c00d46f065e4f6f99ad3e3920769b82 |
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| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T09:11:04Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-5c00d46f065e4f6f99ad3e3920769b822023-11-22T06:00:53ZengMDPI AGMolecules1420-30492021-07-012615462610.3390/molecules26154626Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial CellsSanthosh Chandar Maddila0Chandrashekhar Voshavar1Porkizhi Arjunan2Rashmi Prakash Chowath3Hari Krishna Reddy Rachamalla4Balaji Balakrishnan5Poonkuzhali Balasubramanian6Rajkumar Banerjee7Srujan Marepally8Centre for Stem Cell Research (CSCR) (A Unit of inStem, Bengaluru), Christian Medical College Campus, Bagayam, Vellore 632002, TN, IndiaCollege of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32307, USACentre for Stem Cell Research (CSCR) (A Unit of inStem, Bengaluru), Christian Medical College Campus, Bagayam, Vellore 632002, TN, IndiaCentre for Stem Cell Research (CSCR) (A Unit of inStem, Bengaluru), Christian Medical College Campus, Bagayam, Vellore 632002, TN, IndiaChemical Biology Division, CSIR-Indian Institute of Chemical Technology, Hyderabad 560007, TS, IndiaDepartment of Integrative Biology, School of Bio Sciences and Technology, Vellore Institute of Technology, Vellore 632014, TN, IndiaDepartment of Hematology, Christian Medical College (CMC), Vellore 632004, TN, IndiaDepartment of Hematology, Christian Medical College (CMC), Vellore 632004, TN, IndiaCentre for Stem Cell Research (CSCR) (A Unit of inStem, Bengaluru), Christian Medical College Campus, Bagayam, Vellore 632002, TN, IndiaDelivering nucleic acids into the endothelium has great potential in treating vascular diseases. However, endothelial cells, which line the vasculature, are considered as sensitive in nature and hard to transfect. Low transfection efficacies in endothelial cells limit their potential therapeutic applications. Towards improving the transfection efficiency, we made an effort to understand the internalization of lipoplexes into the cells, which is the first and most critical step in nucleic acid transfections. In this study, we demonstrated that the transient modulation of caveolae/lipid rafts mediated endocytosis with the cholesterol-sequestrating agents, nystatin, filipin III, and siRNA against Cav-1, which significantly increased the transfection properties of cationic lipid-(2-hydroxy-<i>N</i>-methyl-<i>N</i>,<i>N</i>-bis(2-tetradecanamidoethyl)ethanaminium chloride), namely, amide liposomes in combination with 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) (AD Liposomes) in liver sinusoidal endothelial cells (SK-Hep1). In particular, nystatin was found to be highly effective with 2–3-fold enhanced transfection efficacy when compared with amide liposomes in combination with Cholesterol (AC), by switching lipoplex internalization predominantly through clathrin-mediated endocytosis and macropinocytosis.https://www.mdpi.com/1420-3049/26/15/4626cationic lipidsclathrin-mediated endocytosiscaveolae-mediated endocytosismacropinocytosistransfectionendothelial cells |
| spellingShingle | Santhosh Chandar Maddila Chandrashekhar Voshavar Porkizhi Arjunan Rashmi Prakash Chowath Hari Krishna Reddy Rachamalla Balaji Balakrishnan Poonkuzhali Balasubramanian Rajkumar Banerjee Srujan Marepally Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells Molecules cationic lipids clathrin-mediated endocytosis caveolae-mediated endocytosis macropinocytosis transfection endothelial cells |
| title | Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells |
| title_full | Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells |
| title_fullStr | Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells |
| title_full_unstemmed | Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells |
| title_short | Cholesterol Sequestration from Caveolae/Lipid Rafts Enhances Cationic Liposome-Mediated Nucleic Acid Delivery into Endothelial Cells |
| title_sort | cholesterol sequestration from caveolae lipid rafts enhances cationic liposome mediated nucleic acid delivery into endothelial cells |
| topic | cationic lipids clathrin-mediated endocytosis caveolae-mediated endocytosis macropinocytosis transfection endothelial cells |
| url | https://www.mdpi.com/1420-3049/26/15/4626 |
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