Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome
Crouzon syndrome is a rare genetic craniofacial malformation caused by heterozygous gain-of-function mutations in the FGFR2 gene. The resulting constitutive activation of the FGFR2 signaling causes the premature osteogenic differentiation of calvarial mesenchymal stromal cells in skull sutures, lead...
Saved in:
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253124003147 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832589977378095104 |
|---|---|
| author | Federica Tiberio Martina Salvati Luca Polito Giada Tisci Alessia Vita Ornella Parolini Luca Massimi Lorena Di Pietro Pierpaolo Ceci Gianpiero Tamburrini Alessandro Arcovito Elisabetta Falvo Wanda Lattanzi |
| author_facet | Federica Tiberio Martina Salvati Luca Polito Giada Tisci Alessia Vita Ornella Parolini Luca Massimi Lorena Di Pietro Pierpaolo Ceci Gianpiero Tamburrini Alessandro Arcovito Elisabetta Falvo Wanda Lattanzi |
| author_sort | Federica Tiberio |
| collection | DOAJ |
| description | Crouzon syndrome is a rare genetic craniofacial malformation caused by heterozygous gain-of-function mutations in the FGFR2 gene. The resulting constitutive activation of the FGFR2 signaling causes the premature osteogenic differentiation of calvarial mesenchymal stromal cells in skull sutures, leading to early suture ossification. Craniectomy is the gold standard treatment, being invasive and burdened by complications. To address these issues, we developed personalized allele-specific (AS) small interfering RNA (siRNA) to knockdown the expression of the FGFR2 mutant allele in Crouzon patient-derived suture cells. The selected therapeutic siRNA mitigated FGFR2 cascade downregulating phosphorylation of FGFR2 (48%) and of its key effector ERK1/2 (77%) as RUNX2 protein levels (34%). This effect was confirmed by the reduced osteogenic commitment and differentiation of treated cells, evidenced by decreased expression of osteogenic marker genes and a 5-fold decrease in mineralized matrix deposition. We developed a highly biocompatible delivery system for siRNAs, based on human recombinant ferritin nanoparticles (NPs), combining cell targeting with improved nucleic acid encapsulation and endosomal escape properties. We demonstrated the ability of these NPs to deliver and release siRNAs within target cells, sustaining their inhibitory and AS effects. Here, we show that ferritin-mediated AS FGFR2 knockdown by siRNA represents a suitable strategy to dampen FGFR2 overactivation in patients’ cells. |
| format | Article |
| id | doaj-art-5ce506076369457291afcadd01f66806 |
| institution | Kabale University |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj-art-5ce506076369457291afcadd01f668062025-01-24T04:44:58ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-03-01361102427Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndromeFederica Tiberio0Martina Salvati1Luca Polito2Giada Tisci3Alessia Vita4Ornella Parolini5Luca Massimi6Lorena Di Pietro7Pierpaolo Ceci8Gianpiero Tamburrini9Alessandro Arcovito10Elisabetta Falvo11Wanda Lattanzi12Dipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy; Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, ItalyDipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyDipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyDepartment of Biochemical Sciences, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy; CNR-National Research Council of Italy, Institute of Molecular Biology and Pathology, P.le Aldo Moro 7, 00185 Rome, ItalyDipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyDipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy; Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, ItalyUnità Operativa Complessa di Neurochirurgia Infantile, Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, Italy; Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyDipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy; Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, ItalyCNR-National Research Council of Italy, Institute of Molecular Biology and Pathology, P.le Aldo Moro 7, 00185 Rome, ItalyUnità Operativa Complessa di Neurochirurgia Infantile, Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, Italy; Dipartimento di Neuroscienze, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyDipartimento di Scienze Biotecnologiche di Base, Cliniche Intensivologiche e Perioperatorie, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, ItalyCNR-National Research Council of Italy, Institute of Molecular Biology and Pathology, P.le Aldo Moro 7, 00185 Rome, Italy; Corresponding author: Elisabetta Falvo, CNR-National Research Council of Italy, Institute of Molecular Biology and Pathology, Via degli Apuli 4, 00185 Rome, Italy.Dipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy; Unità Operativa Complessa di Neurochirurgia Infantile, Fondazione Policlinico Universitario A. Gemelli IRCCS, Largo Agostino Gemelli 1, 00168 Rome, Italy; Corresponding author: Wanda Lattanzi, Dipartimento Scienze della Vita e Sanità Pubblica, Università Cattolica del Sacro Cuore, Largo Francesco Vito 1, 00168 Rome, Italy.Crouzon syndrome is a rare genetic craniofacial malformation caused by heterozygous gain-of-function mutations in the FGFR2 gene. The resulting constitutive activation of the FGFR2 signaling causes the premature osteogenic differentiation of calvarial mesenchymal stromal cells in skull sutures, leading to early suture ossification. Craniectomy is the gold standard treatment, being invasive and burdened by complications. To address these issues, we developed personalized allele-specific (AS) small interfering RNA (siRNA) to knockdown the expression of the FGFR2 mutant allele in Crouzon patient-derived suture cells. The selected therapeutic siRNA mitigated FGFR2 cascade downregulating phosphorylation of FGFR2 (48%) and of its key effector ERK1/2 (77%) as RUNX2 protein levels (34%). This effect was confirmed by the reduced osteogenic commitment and differentiation of treated cells, evidenced by decreased expression of osteogenic marker genes and a 5-fold decrease in mineralized matrix deposition. We developed a highly biocompatible delivery system for siRNAs, based on human recombinant ferritin nanoparticles (NPs), combining cell targeting with improved nucleic acid encapsulation and endosomal escape properties. We demonstrated the ability of these NPs to deliver and release siRNAs within target cells, sustaining their inhibitory and AS effects. Here, we show that ferritin-mediated AS FGFR2 knockdown by siRNA represents a suitable strategy to dampen FGFR2 overactivation in patients’ cells.http://www.sciencedirect.com/science/article/pii/S2162253124003147MT: Oligonucleotides: Therapies and ApplicationscraniosynostosisCrouzon syndromesmall interfering RNAferritin nanoparticlesgene silencing |
| spellingShingle | Federica Tiberio Martina Salvati Luca Polito Giada Tisci Alessia Vita Ornella Parolini Luca Massimi Lorena Di Pietro Pierpaolo Ceci Gianpiero Tamburrini Alessandro Arcovito Elisabetta Falvo Wanda Lattanzi Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome Molecular Therapy: Nucleic Acids MT: Oligonucleotides: Therapies and Applications craniosynostosis Crouzon syndrome small interfering RNA ferritin nanoparticles gene silencing |
| title | Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome |
| title_full | Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome |
| title_fullStr | Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome |
| title_full_unstemmed | Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome |
| title_short | Targeted allele-specific FGFR2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of Crouzon syndrome |
| title_sort | targeted allele specific fgfr2 knockdown via human recombinant ferritin nanoparticles for personalized treatment of crouzon syndrome |
| topic | MT: Oligonucleotides: Therapies and Applications craniosynostosis Crouzon syndrome small interfering RNA ferritin nanoparticles gene silencing |
| url | http://www.sciencedirect.com/science/article/pii/S2162253124003147 |
| work_keys_str_mv | AT federicatiberio targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT martinasalvati targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT lucapolito targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT giadatisci targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT alessiavita targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT ornellaparolini targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT lucamassimi targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT lorenadipietro targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT pierpaoloceci targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT gianpierotamburrini targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT alessandroarcovito targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT elisabettafalvo targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome AT wandalattanzi targetedallelespecificfgfr2knockdownviahumanrecombinantferritinnanoparticlesforpersonalizedtreatmentofcrouzonsyndrome |