The safety and efficacy of several life-saving protein therapeutics are compromised by unwanted immune responses. Currently there are no viable clinical options to reduce or reverse these responses. Our goal is to develop protein delivery strategies that improve therapeutic efficacy of protein drugs by reducing or reversing unwanted immune responses. During the previous project period, we observed that exposure of mice to protein therapeutics in the presence of phosphatidylserine (PS) resulted in an antigen specific immunological tolerance/hypo-responsiveness towards the protein, leading to our central hypothesis that PS is immune regulatory and has the capacity to convert immunogenic proteins into tolerogens. To fully realize the clinical potential of this observation, it is necessary to determine the structural characteristics of PS that contribute to its ability to convert an immunogen into a tolerogen (SA1), and to use this information to reduce (SA2) and reverse (SA3) unwanted immune responses to therapeutic proteins and improve their therapeutic efficacy. In SA1 we hypothesize that PS converts dendritic cells from an immunogenic to a tolerogenic phenotype, generating regulatory T-cells that in turn suppress antigen-specific T-cells and B-cells. We investigate the structural and biophysical determinants of PS that program dendritic cells to send tolerogenic over immunogenic signals. Since pre-exposure of mice to FVIII in the presence of PS significantly reduced their immune response during re-challenge with FVIII, in SA2, we investigate a similar pre-treatment approach with two other immunogenic therapeutic proteins to prevent or reduce unwanted immune responses to these proteins during therapy. Specifically we will test this approach with acid alpha glucosidase (GAA in Pompe disease, a lysosomal storage disorder) and Factor IX (in Hemophilia B, a bleeding disorder). We will determine the immune cells that are involved in the induction of tolerance, and establish the impact of such tolerance induction on the efficacy of these two replacement therapies. In SA3, we develop clinical strategies to reverse established responses to FVIII, FIX and GAA, and thereby improve the efficacy of treatments for these bleeding and lysosomal disorders. Since PS is exposed in outer layer of secreted vesicles such as apoptotic bodies and on exosomes released from cells, we propose and will test the hypothesis that lipid particles mimicking these natural vesicles will reverse the established immune responses to therapeutic proteins. Our goal in SA3 is the optimization of immunization protocol and testing of an artificial lipid particle tha mimics the secreted vesicles, to reverse the unwanted established immune responses. These studies are expected to lead to novel clinical strategies to reduce and reverse immune responses to therapeutic proteins and thereby improve the efficacy of several lifesaving protein-based therapies.

Public Health Relevance

Development of unwanted immune response against therapeutic proteins is a major clinical complication that impacts safety and efficacy of several life- saving protein based therapies. Once such responses are established, either no alternate therapies are available or cost prohibitive therapies are attempted with limited success. The proposed research is relevant to public health because the proposal is focused on developing novel clinical approaches to reduce and reverse such unwanted immune responses and is relevant to the part of NIH's mission of seeking fundamental knowledge to enhance human health.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL070227-15
Application #
9394031
Study Section
Xenobiotic and Nutrient Disposition and Action Study Section (XNDA)
Program Officer
Sarkar, Rita
Project Start
2002-04-01
Project End
2019-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
15
Fiscal Year
2018
Total Cost
Indirect Cost
Name
State University of New York at Buffalo
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
038633251
City
Amherst
State
NY
Country
United States
Zip Code
14228
Schneider, Jennifer L; Dingman, Robert K; Balu-Iyer, Sathy V (2017) Lipidic Nanoparticles Comprising Phosphatidylinositol Mitigate Immunogenicity and Improve Efficacy of Recombinant Human Acid Alpha-Glucosidase in a Murine Model of Pompe Disease. J Pharm Sci :
Ramakrishnan, Radha; Balu-Iyer, Sathy V (2016) Effect of Biophysical Properties of Phosphatidylserine Particle on Immune Tolerance Induction Toward Factor VIII in a Hemophilia A Mouse Model. J Pharm Sci 105:3039-3045
Shetty, Krithika A; Kosloski, Matthew P; Mager, Donald E et al. (2016) Factor VIII associated with lipidic nanoparticles retains efficacy in the presence of anti-factor VIII antibodies in hemophilia A mice. Biopharm Drug Dispos 37:409-420
Schneider, Jennifer L; Balu-Iyer, Sathy V (2016) Phosphatidylserine Converts Immunogenic Recombinant Human Acid Alpha-Glucosidase to a Tolerogenic Form in a Mouse Model of Pompe Disease. J Pharm Sci 105:3097-3104
Shetty, Krithika A; Merricks, Elizabeth P; Raymer, Robin et al. (2016) Soy Phosphatidylinositol-Containing Lipid Nanoparticle Prolongs the Plasma Survival and Hemostatic Efficacy of B-domain-Deleted Recombinant Canine Factor VIII in Hemophilia A Dogs. J Pharm Sci 105:2459-64
Ramakrishnan, Radha; Davidowitz, Andrew; Balu-Iyer, Sathy V (2015) Exposure of FVIII in the Presence of Phosphatidyl Serine Reduces Generation of Memory B-Cells and Induces Regulatory T-Cell-Mediated Hyporesponsiveness in Hemophilia A Mice. J Pharm Sci 104:2451-6
Gaitonde, Puneet; Purohit, Vivek S; Balu-Iyer, Sathy V (2015) Intravenous administration of Factor VIII-O-Phospho-L-Serine (OPLS) complex reduces immunogenicity and preserves pharmacokinetics of the therapeutic protein. Eur J Pharm Sci 66:157-62
Fathallah, Anas M; Balu-Iyer, Sathy V (2015) Anatomical, physiological, and experimental factors affecting the bioavailability of sc-administered large biotherapeutics. J Pharm Sci 104:301-6
Kelleher Jr, Raymond J; Balu-Iyer, Sathy; Loyall, Jenni et al. (2015) Extracellular Vesicles Present in Human Ovarian Tumor Microenvironments Induce a Phosphatidylserine-Dependent Arrest in the T-cell Signaling Cascade. Cancer Immunol Res 3:1269-78
Fathallah, Anas M; Turner, Michael R; Mager, Donald E et al. (2015) Effects of hypertonic buffer composition on lymph node uptake and bioavailability of rituximab, after subcutaneous administration. Biopharm Drug Dispos 36:115-25

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