Type 1 diabetes (T1D) develops as a result of insufficient insulin being produced due to a self-destructive immune response against insulin producing beta cells. Although a number of factors are known to promote advantageous immune cell responses in experimental systems for T1D, systemic intravenous delivery of these agents often results in significant harmful off-target effects due to the uncontrolled dosing of bystander cells, tissues and organs. This project focuses on the targeted in vivo delivery of pro-tolerance factors and insulin antigen, in particulate form, targeted to a key immune cell type, dendritic cells (DCs). Dendritic cells, critical for maintenance and initiation of immunity to foregn antigens and tolerance to self-antigens, are phagocytic, antigen presenting cells. This makes DCs an ideal recipient for the targeted delivery of agents provided in particulate form. Moreover, exogenous conditioning of DCs with certain immunomodulatory agents has been shown to induce a pro-tolerance DC phenotype as well as ameliorate T1D. Vaccination with DC-targeting microparticles (MPs) holds promise to correct T1D autoimmune responses, critically, without the costly ex vivo manipulations required of DC-based cellular therapy. This enables the potential for widespread use. Micron-sized biodegradable polymeric particles are phagocytosable, which effectively promotes delivery of encapsulated factors to intracellular sites of DCs over non-phagocytes. These phagocytosable particles can be further targeted to DCs by surface immobilizing molecules targeting DC receptors. Larger (but still small enough to be injectable), non-phagocytosable biodegradable polymeric particles provide controlled release of encapsulated factors to the local extracellular environment at the subcutaneous injection site. Encapsulated factors in these large particles consist of bioactive factors for which DCs have the cognate cell-surface receptors. The objective of this proposal is to engineer a subcutaneously injectable dual MP vaccine system consisting of i.) Phagocytosable DC-targeting MPs delivering antigen and immunomodulatory factor (insulin and vitamin D3) to intracellular sites;and ii.) Non-phagocytosable MPs to deliver, extracellular, factors (GM- CSF and TGF-b1) for DC recruitment and tolerance induction. We expect to effect a pro-tolerogenic DC phenotype and promote induction of regulatory T-cells, suppression of auto-reactive T-cells, and prevent and reverse diabetes in non-obese diabetic (NOD) mice. We hypothesize that the combination of the multiple components in the dual MP system will more effectively provide robust, durable antigen-specific immune suppression than single-component formulations, either in MPs or in soluble form.
Aim 1 is to formulate the dual MP system, test it in vitro by characterizing DC phenotype (activated, immature or tolerogenic) and T-cell response (stimulation, Th1, Th2, Treg, or Th17).
Aim 2 is to evaluate the ability of the dual MP formulation in vivo, aiming to prevent and reverse diabetes in NOD mice. This novel and innovative approach holds promise for correcting autoimmune responses in T1D and represents a simple, clinically translatable system.

Public Health Relevance

Type-1 diabetes is an autoimmune disease with significant personal and economic impact in the US. Therapeutic vaccination approaches for type-1 diabetes hold promise to correct these antigen-specific autoimmune responses. The objective of this proposal is to engineer polymeric biomaterials-based microparticles as an injectable vaccine system to retrain the immune system to correct aberrant activation toward self antigens.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK091658-03
Application #
8636464
Study Section
Biomaterials and Biointerfaces Study Section (BMBI)
Program Officer
Pawlyk, Aaron Christopher
Project Start
2012-04-01
Project End
2017-03-31
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
3
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Florida
Department
Biomedical Engineering
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Fernando, Lawrence P; Lewis, Jamal S; Evans, Brian C et al. (2017) Formulation and characterization of poly(propylacrylic acid)/poly(lactic-co-glycolic acid) blend microparticles for pH-dependent membrane disruption and cytosolic delivery. J Biomed Mater Res A :
Yang, Lirong; Bracho-Sanchez, Evelyn; Fernando, Lawrence P et al. (2017) Poly(2-propylacrylic acid)/poly(lactic-co-glycolic acid) blend microparticles as a targeted antigen delivery system to direct either CD4+ or CD8+ T cell activation. Bioeng Transl Med 2:202-211
Cho, Jonathan J; Stewart, Joshua M; Drashansky, Theodore T et al. (2017) An antigen-specific semi-therapeutic treatment with local delivery of tolerogenic factors through a dual-sized microparticle system blocks experimental autoimmune encephalomyelitis. Biomaterials 143:79-92
Zaveri, Toral D; Dolgova, Natalia V; Lewis, Jamal S et al. (2017) Macrophage integrins modulate response to ultra-high molecular weight polyethylene particles and direct particle-induced osteolysis. Biomaterials 115:128-140
Stewart, Joshua M; Keselowsky, Benjamin G (2017) Combinatorial drug delivery approaches for immunomodulation. Adv Drug Deliv Rev 114:161-174
Keselowsky, Benjamin G; Lewis, Jamal S (2017) Dendritic cells in the host response to implanted materials. Semin Immunol 29:33-40
Bracho-Sanchez, E; Xia, C Q; Clare-Salzler, M J et al. (2016) Micro and Nano Material Carriers for Immunomodulation. Am J Transplant 16:3362-3370
Acharya, Abhinav P; Carstens, Matthew R; Lewis, Jamal S et al. (2016) A cell-based microarray to investigate combinatorial effects of microparticle-encapsulated adjuvants on dendritic cell activation. J Mater Chem B 4:1672-1685
Lewis, Jamal S; Dolgova, Natalia V; Zhang, Ying et al. (2015) A combination dual-sized microparticle system modulates dendritic cells and prevents type 1 diabetes in prediabetic NOD mice. Clin Immunol 160:90-102
Carstens, Matthew R; Fisher, Robert C; Acharya, Abhinav P et al. (2015) Drug-eluting microarrays to identify effective chemotherapeutic combinations targeting patient-derived cancer stem cells. Proc Natl Acad Sci U S A 112:8732-7

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