Therapeutic effect of the immunomodulatory drug lenalidomide, but not pomalidomide, in experimental models of rheumatoid arthritis and inflammatory bowel disease

Abstract

Thalidomide is an immunomodulatory drug (IMiD) with proven therapeutic action in several autoimmune/inflammatory diseases; however, its inherent high toxicity has led to the development of more powerful and safer thalidomide analogs, including lenalidomide and pomalidomide. These are new generation IMiDs that exhibit direct antitumor activity as well as anti-inflammatory/immunomodulatory properties, and are FDA-approved for the treatment of several hematological malignances. Here we investigated the potential therapeutic effects of lenalidomide and pomalidomide in several experimental murine models of autoimmune/inflammatory diseases: 2,4,6-trinitrobenzene sulfonic acid- and dextran sulfate sodium-induced inflammatory bowel disease and type II collagen-induced arthritis. Lenalidomide displayed a strong therapeutic effect in all these models of autoimmune/inflammatory diseases, while the effect of pomalidomide was less pronounced. In vitro experiments confirmed the immunosuppressive effect of both IMiDs on the proliferative response of stimulated human lymphocytes and on the balance of secreted cytokines toward an anti-inflammatory profile. We conclude that lenalidomide may offer a therapeutic opportunity against autoimmune/inflammatory diseases.

Figure 1

Lenalidomide treatment protects against 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced acute severe colitis. Colitis was induced by intracolonic administration of TNBS and mice were treated i.p. with lenalidomide or pomalidomide for 3 days, starting at 12 h after TNBS injection. Clinical evolution was monitored by determining the daily body weight loss and survival (a) as well as measuring the colitis score (at day 3) (b), the macroscopic colonic damage score (c) and the histopathologic score (at day 3) (d). Data are expressed as mean±s.e.m.; n=10 mice per group in a–c; n=5 mice per group in d. Statistical differences between groups were calculated as described in Materials and methods. *P<0.05; ***P<0.01; ****P<0.001.

Figure 2

Lenalidomide treatment protects from dextran sulfate sodium (DSS)-induced acute colitis. Mice received 3% DSS in the drinking water from day 0 to day 7. Lenalidomide or pomalidomide were administered intraperitoneally once a day for 3 days, starting at day 3. (a) Disease activity scores were determined daily. (b) Colon damage scores and colon length and weight were determined at day 8. Mice receiving tap water instead of DSS were used as controls (naive). Data are expressed as mean±s.e.m.; n=10 mice per group in a; n=5 mice per group in b. Statistical differences between groups were calculated as described in Materials and methods. *P<0.05; ^**P<0.01.

Figure 3

Lenalidomide treatment ameliorates established collagen-induced arthritis (CIA). Arthritis score (a) and number of affected paws (b) after therapeutic administration of lenalidomide and pomalidomide in CIA mice. (c) Serum levels of IL6, IL1α, IL17, IL13, IL4 and IL27 in control- and lenalidomide-treated arthritic mice. (d) Levels of anti-CII antibodies in sera from control and lenalidomide-treated mice. Data pooled from two independent experiments with eight mice per experimental group (n=16 mice per group). Mean±s.e.m. of experimental groups is shown in a and b, and values from individual mice and the mean of experimental groups are shown in c and d. Statistical differences between groups were calculated as described in Materials and methods. *P<0.05; **P<0.01; ***P<0.001. RU, relative units.

Figure 4

In vitro immunosuppressive and anti-inflammatory properties of lenaledomide and pomaledomide. (a) Carboxyfluorescein diacetate succinimidyl ester (CSFE)-labeled peripheral blood mononuclear cells (PBMCs) from healthy donors were stimulated or not with phytohemaglutinin (PHA). Then, PHA-stimulated PBMCs were treated with or without 10 μM of lenalidomide or pomalidomide for 5 days. The number of cycling (CSFE^mild/low) cells was determined by flow cytometry. Upper panel: percentage of non-proliferating cells expressed as mean±s.e.m. Lower panel: representative flow cytometry histograms of CSFE-labeled PBMCs. R1, proliferating cells; R2 non-proliferating cells. (b) Cytokine concentrations in cell culture supernatants were determined by Luminex Multiplex assays. PBMCs from healthy donors were stimulated or not with lipopolysaccharide (LPS or PHA for interferon gamma (IFNγ), IL17 and IL13). Stimulated PBMCs were treated with or without 10 μM of lenalidomide or pomalidomide. Data are expressed as mean±s.e.m. Statistical differences between groups were calculated as described in Materials and methods. *P<0.05; **P<0.01; RU, relative units.