Oral histone deacetylase inhibitor synergises with T cell targeted immunotherapy to preserve beta cell metabolic function and induce stable remission of new-onset autoimmune diabetes in NOD mice
Combination therapy targeting the major actors involved in the immune-mediated destruction of pancreatic beta cells appears to be an indispensable approach to treat type 1 diabetes effectively. We hypothesised that the combination of an orally active pan-histone deacetylase inhibitor (HDACi: givinostat) with subtherapeutic doses of CD3 antibodies may provide ideal synergy to treat ongoing autoimmunity.
NOD mice transgenic for the human CD3ε (also known as CD3E) chain (NOD-huCD3ε) were treated for recent-onset diabetes with oral givinostat, subtherapeutic doses of humanised CD3 antibodies (otelixizumab, 50 μg/day, 5 days, i.v.) or a combination of both drugs. Disease remission, metabolic profiles and autoreactive T cell responses were analysed in treated mice.
We demonstrated that givinostat synergised with otelixizumab to induce durable remission of diabetes in 80% of recently diabetic NOD-huCD3ε mice. Remission was obtained in only 47% of mice treated with otelixizumab alone. Oral givinostat monotherapy did not reverse established diabetes but reduced the in situ production of inflammatory cytokines (IL-1β, IL-6, TNF-α). Importantly, the otelixizumab + givinostat combination strongly improved the metabolic status of NOD-huCD3ε mice; the mice recovered the capacity to appropriately produce insulin, control hyperglycaemia and sustain glucose tolerance. Finally, diabetes remission induced by the combination therapy was associated with a significant reduction of insulitis and autoantigen-specific CD8+ T cell responses.
HDACi and low-dose CD3 antibodies synergised to abrogate in situ inflammation and thereby improved pancreatic beta cell survival and metabolic function leading to long-lasting diabetes remission. These results support the therapeutic potential of protocols combining these two drugs, both in clinical development, to restore self-tolerance and insulin independence in type 1 diabetes.
KeywordsBeta cells Glucose tolerance HDACi Human CD3 antibodies Humanised NOD mice Insulin secretion Type 1 diabetes
Forkhead box P3
Histone deacetylase inhibitor
Islet-specific glucose-6-phosphatase catalytic subunit related protein
Lymphocyte-activation gene 3
Pancreatic lymph node
Regulatory type 1 cell
Regulatory T cell
The authors thank M. Bellanger (INSERM U1151, Paris, France) for taking care of the NOD-huCD3ε mouse colony and for providing technical assistance for the experimental mouse work. We are also grateful to E. Panafieu, S. Fonlebeck and Y. Loudin (INSERM U1151, Department of Immunology, Paris, France) for mouse production and maintenance.
The data generated during the current study are available from the corresponding author on reasonable request.
This work was supported by grants from the JDRF (#1-2011-654), institutional funding from INSERM and University Paris Descartes and also with the support of Fondation Day Solvay and Fondation Centaure. AB was supported by a doctoral fellowship from INSERM and by a grant from the Société Française d’Endocrinologie et Diabétologie Pédiatrique (grant from Novo Nordisk). The funders had no role in study design, data collection, interpretation or decision to submit the work for publication.
Duality of interest
The authors declare that there is no duality of interest associated with this manuscript.
AB designed experiments, acquired and analysed data, and wrote the manuscript. TG and FV designed and performed experiments, and analysed data. MSD provided research material and contributed to the design of the experiments. LC provided critical advice and help in writing the manuscript. TM-P initiated the study with LC and contributed to planning the protocol and reviewed the manuscript. All authors revised the manuscript and approved the final version to be published. SY designed and directed the study, analysed the data and wrote the manuscript. SY is the guarantor of this work.
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