Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity
<b>Background:</b> The C1858T <i>PTPN22</i> variant is strongly associated with type 1 diabetes and autoimmune thyroid disease. Current treatment is substitutive hormonal administration, which does not target the disease pathogenetic mechanism. We previously implemented a nov...
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2025-05-01
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| author | Simona Sennato Giorgia Paldino Cecilia Bombelli Irene Mezzani Stefania Petrini Domenico Vittorio Delfino Francesco Fiorentino Carlotta Marianecci Alessia Ciogli Dante Rotili Francesca Ceccacci Alessandra Fierabracci |
| author_facet | Simona Sennato Giorgia Paldino Cecilia Bombelli Irene Mezzani Stefania Petrini Domenico Vittorio Delfino Francesco Fiorentino Carlotta Marianecci Alessia Ciogli Dante Rotili Francesca Ceccacci Alessandra Fierabracci |
| author_sort | Simona Sennato |
| collection | DOAJ |
| description | <b>Background:</b> The C1858T <i>PTPN22</i> variant is strongly associated with type 1 diabetes and autoimmune thyroid disease. Current treatment is substitutive hormonal administration, which does not target the disease pathogenetic mechanism. We previously implemented a novel immunotherapy, employing siRNA directed against the C1858T variant of <i>PTPN22</i> delivered via functionalized lipoplexes, in order to halt autoimmune disease progression. <b>Objectives</b>: The objective of this study was to optimize lipoplex formulations functionalized with F9-PEG (a Siglec-10’s ligand) to facilitate targeted delivery by investigating their physical and chemical properties to warrant the best performance in in vivo studies. <b>Methods</b>: The effectiveness of siRNA liposome binding was evaluated by varying the relative lipid/siRNA charge ratio and analyzing the stability of the different formulations with respect to the methods of F9-PEG addition and ATTO740 fluorescent labeling by electrophoresis, dynamic and dielectrophoretic light scattering (DLS and DELS), and high-performance liquid chromatography (HPLC). <b>Results</b>: The optimal charge ratio of +2/−1 (lipid/siRNA) ensured a greater stability of lipoplexes and complete complexation of siRNA. Stability was improved by selecting a protocol of preparation that envisages functionalization with F9-PEG and the addition of ATTO740 lipid in the lipid film preparation step. HPLC confirmed the integrity of siRNA after preparation. <b>Conclusions</b>: The results of this study lead to formulations of F9-PEG lipoplexes with optimal properties that could be used for biodistribution and safety/efficacy studies in mice. Lipoplexes functionalized with F9-PEG could therefore represent a promising personalized nanotherapeutic platform for targeted siRNA delivery in endocrine C1858T patients. |
| format | Article |
| id | doaj-art-5b2f8227c30a42489bd0aaaed25fcad8 |
| institution | Kabale University |
| issn | 1999-4923 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
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| series | Pharmaceutics |
| spelling | doaj-art-5b2f8227c30a42489bd0aaaed25fcad82025-08-20T03:29:40ZengMDPI AGPharmaceutics1999-49232025-05-0117671010.3390/pharmaceutics17060710Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine AutoimmunitySimona Sennato0Giorgia Paldino1Cecilia Bombelli2Irene Mezzani3Stefania Petrini4Domenico Vittorio Delfino5Francesco Fiorentino6Carlotta Marianecci7Alessia Ciogli8Dante Rotili9Francesca Ceccacci10Alessandra Fierabracci11CNR-Institute for Complex Systems (ISC), and Physics Department, Sapienza University of Rome, P. le A. Moro 5, 00185 Rome, ItalyBambino Gesù Children’s Hospital, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), 00146 Rome, ItalyCNR-Institute for Biological Systems, Secondary Office of Rome—Sapienza University of Rome, 00185 Rome, ItalyBambino Gesù Children’s Hospital, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), 00146 Rome, ItalyBambino Gesù Children’s Hospital, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), 00146 Rome, ItalyDepartment of Medicine and Surgery, University of Perugia, 06129 Perugia, ItalyDepartment of Drug Chemistry and Technologies, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Drug Chemistry and Technologies, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Drug Chemistry and Technologies, Sapienza University of Rome, 00185 Rome, ItalyDepartment of Science, Roma Tre University, 00146 Rome, ItalyCNR-Institute for Biological Systems, Secondary Office of Rome—Sapienza University of Rome, 00185 Rome, ItalyBambino Gesù Children’s Hospital, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS), 00146 Rome, Italy<b>Background:</b> The C1858T <i>PTPN22</i> variant is strongly associated with type 1 diabetes and autoimmune thyroid disease. Current treatment is substitutive hormonal administration, which does not target the disease pathogenetic mechanism. We previously implemented a novel immunotherapy, employing siRNA directed against the C1858T variant of <i>PTPN22</i> delivered via functionalized lipoplexes, in order to halt autoimmune disease progression. <b>Objectives</b>: The objective of this study was to optimize lipoplex formulations functionalized with F9-PEG (a Siglec-10’s ligand) to facilitate targeted delivery by investigating their physical and chemical properties to warrant the best performance in in vivo studies. <b>Methods</b>: The effectiveness of siRNA liposome binding was evaluated by varying the relative lipid/siRNA charge ratio and analyzing the stability of the different formulations with respect to the methods of F9-PEG addition and ATTO740 fluorescent labeling by electrophoresis, dynamic and dielectrophoretic light scattering (DLS and DELS), and high-performance liquid chromatography (HPLC). <b>Results</b>: The optimal charge ratio of +2/−1 (lipid/siRNA) ensured a greater stability of lipoplexes and complete complexation of siRNA. Stability was improved by selecting a protocol of preparation that envisages functionalization with F9-PEG and the addition of ATTO740 lipid in the lipid film preparation step. HPLC confirmed the integrity of siRNA after preparation. <b>Conclusions</b>: The results of this study lead to formulations of F9-PEG lipoplexes with optimal properties that could be used for biodistribution and safety/efficacy studies in mice. Lipoplexes functionalized with F9-PEG could therefore represent a promising personalized nanotherapeutic platform for targeted siRNA delivery in endocrine C1858T patients.https://www.mdpi.com/1999-4923/17/6/710type 1 diabetesendocrine autoimmunityimmunotherapyvariant <i>PTPN22</i>lipoplexes optimization |
| spellingShingle | Simona Sennato Giorgia Paldino Cecilia Bombelli Irene Mezzani Stefania Petrini Domenico Vittorio Delfino Francesco Fiorentino Carlotta Marianecci Alessia Ciogli Dante Rotili Francesca Ceccacci Alessandra Fierabracci Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity Pharmaceutics type 1 diabetes endocrine autoimmunity immunotherapy variant <i>PTPN22</i> lipoplexes optimization |
| title | Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity |
| title_full | Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity |
| title_fullStr | Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity |
| title_full_unstemmed | Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity |
| title_short | Optimization of Lipoplexes Functionalized with a Sialic Acid Mimetic (F9-PEG) to Target the C1858T <i>PTPN22</i> Variant for Preclinical Assessment of a Novel Immunotherapy in Endocrine Autoimmunity |
| title_sort | optimization of lipoplexes functionalized with a sialic acid mimetic f9 peg to target the c1858t i ptpn22 i variant for preclinical assessment of a novel immunotherapy in endocrine autoimmunity |
| topic | type 1 diabetes endocrine autoimmunity immunotherapy variant <i>PTPN22</i> lipoplexes optimization |
| url | https://www.mdpi.com/1999-4923/17/6/710 |
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