Your browser doesn't support javascript.
loading
Aptabinding of tumor necrosis factor-α (TNFα) inhibits its proinflammatory effects and alleviates islet inflammation.
Mahmoud, Roaa; Kalivarathan, Jagan; Castillo, Abdul J; Wang, Sasha; Fuglestad, Brian; Kanak, Mazhar A; Dhakal, Soma.
Afiliación
  • Mahmoud R; Department of Chemistry, Virginia Commonwealth University, Richmond, Virginia, USA.
  • Kalivarathan J; Department of Surgery, Virginia Commonwealth University - School of Medicine, Virginia, USA.
  • Castillo AJ; Islet Cell Lab, Hume-Lee Transplant Center, VCU Health System, Richmond, Virginia, USA.
  • Wang S; Department of Chemistry, Virginia Commonwealth University, Richmond, Virginia, USA.
  • Fuglestad B; Department of Chemistry, Virginia Commonwealth University, Richmond, Virginia, USA.
  • Kanak MA; Department of Chemistry, Virginia Commonwealth University, Richmond, Virginia, USA.
  • Dhakal S; Institute for Structural Biology, Drug Discovery and Development, Virginia Commonwealth University, Richmond, Virginia, USA.
Biotechnol J ; 19(1): e2300374, 2024 Jan.
Article en En | MEDLINE | ID: mdl-37772688
Pancreatic islet cell transplantation (ICT) has emerged as an effective therapy for diabetic patients lacking endogenous insulin production. However, the islet graft function is compromised by a nonspecific inflammatory and thrombotic reaction known as the instant blood-meditated inflammatory reaction (IBMIR). Here, we report the characterization of four single-stranded DNA aptamers that bind specifically to TNFα - a pivotal cytokine that causes proinflammatory signaling during the IBMIR process - using single molecule binding analysis and functional assays as a means to assess the aptamers' ability to block TNFα activity and inhibiting the downstream proinflammatory gene expression in the islets. Our single-molecule fluorescence analyses of mono- and multivalent aptamers showed that they were able to bind effectively to TNFα with monoApt2 exhibiting the strongest binding (Kd  âˆ¼ 0.02 ± 0.01 nM), which is ∼3 orders of magnitude smaller than the Kd of the other aptamers. Furthermore, the in vitro cell viability analysis demonstrated an optimal and safe dosage of 100 µM for monoApt2 compared to 50 µM for monoApt1 and significant protection from proinflammatory cytokine-mediated cell death. More interestingly, monoApt2 reversed the upregulation of IBMIR mediating genes induced by TNFα in the human islets, and this was comparable to established TNFα antagonists. Both monoaptamers showed high specificity and selectivity for TNFα. Collectively, these findings suggest the potential use of aptamers as anti-inflammatory and localized immune-modulating agents for cellular transplant therapy.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trasplante de Islotes Pancreáticos / Factor de Necrosis Tumoral alfa Límite: Humans Idioma: En Revista: Biotechnol J Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Alemania
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Trasplante de Islotes Pancreáticos / Factor de Necrosis Tumoral alfa Límite: Humans Idioma: En Revista: Biotechnol J Asunto de la revista: BIOTECNOLOGIA Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Alemania