Granulocyte-Colony Stimulating Factor-Overexpressing Mesenchymal Stem Cells Exhibit Enhanced Immunomodulatory Actions Through the Recruitment of Suppressor Cells in Experimental Chagas Disease Cardiomyopathy

Abstract

Genetic modification of mesenchymal stem cells (MSCs) is a promising strategy to improve their therapeutic effects. Granulocyte-colony stimulating factor (G-CSF) is a growth factor widely used in the clinical practice with known regenerative and immunomodulatory actions, including the mobilization of regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). Here we evaluated the therapeutic potential of MSCs overexpressing G-CSF (MSC_G-CSF) in a model of inflammatory cardiomyopathy due to chronic Chagas disease. C57BL/6 mice were treated with wild-type MSCs, MSC_G-CSF, or vehicle (saline) 6 months after infection with Trypanosoma cruzi. Transplantation of MSC_G-CSF caused an increase in the number of circulating leukocytes compared to wild-type MSCs. Moreover, G-CSF overexpression caused an increase in migration capacity of MSCs to the hearts of infected mice. Transplantation of either MSCs or MSC_G-CSF improved exercise capacity, when compared to saline-treated chagasic mice. MSC_G-CSF mice, however, were more potent than MSCs in reducing the number of infiltrating leukocytes and fibrosis in the heart. Similarly, MSC_G-CSF-treated mice presented significantly lower levels of inflammatory mediators, such as IFNγ, TNFα, and Tbet, with increased IL-10 production. A marked increase in the percentage of Tregs and MDSCs in the hearts of infected mice was seen after administration of MSC_G-CSF, but not MSCs. Moreover, Tregs were positive for IL-10 in the hearts of T. cruzi-infected mice. In vitro analysis showed that recombinant hG-CSF and conditioned medium of MSC_G-CSF, but not wild-type MSCs, induce chemoattraction of MDSCs in a transwell assay. Finally, MDSCs purified from hearts of MSC_G-CSF transplanted mice inhibited the proliferation of activated splenocytes in a co-culture assay. Our results demonstrate that G-CSF overexpression by MSCs potentiates their immunomodulatory effects in our model of Chagas disease and suggest that mobilization of suppressor cell populations such as Tregs and MDSCs as a promising strategy for the treatment of chronic Chagas disease. Finally, our results reinforce the therapeutic potential of genetic modification of MSCs, aiming at increasing their paracrine actions.

Strategies to Potentiate Paracrine Therapeutic Efficacy of Mesenchymal Stem Cells in Inflammatory Diseases

“…In addition, IL-10, another important anti-inflammatory cytokine, IL-4, can be genetically delivered in MSCs to enhance the immunosuppressive role of naive cells targeting autoimmune disorders [ 103 , 104 ]. To increase homing capacity to the injury site, Silva et al introduced granulocyte-Colony Stimulating Factor (GM-CSF) to MSCs (MSC GM-CSF ) and evaluated their therapeutic roles in Chagas disease cardiomyopathy [ 105 ]. Compared to control MSCs, MSC GM-CSF displayed a remarkable homing ability to the heart, sequentially leading to the recruitment of myeloid-derived suppressor cells (MDSCs) and Treg induction.…”

Genetic Engineering as a Strategy to Improve the Therapeutic Efficacy of Mesenchymal Stem/Stromal Cells in Regenerative Medicine

“…Lentiviruses, however, have the property of transducing dividing and quiescent cells (Lewis et al, 1992;Vargas et al, 2016). Currently, lentiviral vectors are Noiseux et al, 2006 Rabbit amniotic fluid Lentivirus Heart ischemia-reperfusion injury Wang et al, 2016 Human umbilical cord Adenovirus Acute myocardial infarction Ma et al, 2017 Kopru et al, 2018 Glycogen synthase kinase-3β (GSK-3β) Mouse bone marrow Adenovirus Myocardial infarction Cho et al, 2011 Granulocyte chemotactic protein (GCP)-2 Human adipose tissue Lentivirus Myocardial infarction Kim et al, 2012 Granulocyte-colony stimulating factor (G-CSF) Mouse bone marrow Lentivirus Chagas disease cardiomyopathy Silva et al, 2018b Heme oxygenase-1 (HO-1) Mouse adipose tissue Plasmid transfection Heart ischemic injury Preda et al, 2014 Rat bone marrow Lentivirus Acute lung injury Dog adipose tissue Lentivirus Spinal cord injury Lee et al, 2017 Human embryonic stem cell…”

“…The therapeutic effects of MSCs overexpression growth factors were also studied in a mouse model of cardiomyopathy induced by chronic infection with Trypanosoma cruzi , the causative agent of Chagas disease. Administration of genetically modified mesenchymal cells overexpressing granulocyte colony-stimulating factor (G-CSF) were about 2-fold more potent in reducing the number of inflammatory cells and the percentage of fibrosis than control MSCs, acting by increasing the number of myeloid-derived suppressor cells and T regulatory cells ( Silva et al, 2018b ). In contrast, no increase in anti-inflammatory or antifibrotic activity was seen after transplantation of IGF-overexpressing MSCs in T. cruzi -infected animals.…”

Uso de Células Tronco Mesenquimais na Medicina Regenerativa e Rejuvenescimento

“…O que abre caminho para novos tratamentos, podendo ser pelo transplante de células ou pela modificação genética de células de forma que isso aumente a sua ação parácrina, reduzindo a reação imune na Doença de Chagas. 27,28 O mesmo grupo, juntamente com o Dr. Gilson Soares Feitosa e colaboradores, desenvolveram pesquisa inédita no Brasil em pacientes do Hospital Santa Izabel, demonstrando que a injeção intracoronariana de um aspirado processado da medula óssea promoveu resultados positivos no tratamento de um paciente vítima de insuficiência cardíaca secundária à doença de Chagas. Nesse caso, a fração de ejeção do ventrículo esquerdo (FEVE) em repouso aumentou de 24% para 32% depois de 30 dias do procedimento, sem alterar o esquema medicamentoso.…”