Terapia gênica com o fator estimulante de colônias de macrófagos para isquemia de membro em modelo murino

• Ano de defesa: 2017
• Autor(a) principal: Gallo, Camila Congentino [UNIFESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Gene therapy; Ischaemia; Musculoskeletal system; Macrophages; Factor stimulating macrophage colony; Terapia gênica; Isquemia; Sistema musculoesquelético; Macrófagos; Fator estimulante de colônia de macrófagos
• Link de acesso: https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=5015433; http://repositorio.unifesp.br/handle/11600/50526

Background: Peripheral artery disease is characterized by the narrowing or occlusion of arteries that supply the limbs, possibly evolving to chronic critical ischemia. M-CSF promotes proliferation, differentiation and survival of monocytes and macrophages, which are essential elements for vessel formation. M-CSF also differentiates preferentially monocytes to M2 macrophages, which are antiinflammatory and produce angiogenic factors. Aims: To assess the effect of M-CSF gene therapy on functional restoration of murine ischemic limbs. Methods: Hindlimb ischemia was surgically induced in 10-12 weeks old Balb/c and, three days later, the rectus-femoris muscle was electroporated with 100).1gof uPMCSF plasmid. Temporal gene expression was obtained by ELlSA. Over the month, the animais were monitored by Laser Doppler Perfosion Imaging (LDPI) and their cellular profile was obtained by hemogram. Thirty days after transfection, gastrocnemius muscle force and mass were determined. Histological analyses were processed with gastrocnemius muscles at the end of the month. The following groups were included: Ischemic mice (I), ischemic mice electroporated with empty uP plasmid (luP), ischemic mice electroporated with uP-MCSF (1M) and non-ischemic mice (NI). Results: M-CSF expression was increased on days 2, 4 and 7, reaching baseline on day 14 on the 1Mgroup. The ratio of gastrocnemius force/mass (N/g) was 3.4±0.6 for the NI group, 0.4±0.1 for the I group, 0.4±0.2 for the luP group and 2.3±1.0 for the 1M group, although LDPI didn't show any significant difference in blood flow among groups. Histological analyses showed an increased average perimeter for 1M group in comparison to NI and I groups (140.4±15.8; 101.4±17.2 and 118.3±10.4 um, respectively). Conclusion: M-CSF gene therapy increased vessel caliber, normalized muscle structure and improved muscle functionality.