Heat shock protein 90 inhibitor RGRN-305 potently attenuates skin inflammation

• DOI: 10.3389/fimmu.2023.1128897
• Publication Date: 2023-02-07T09:01:53Z
• Authors: Hakim Ben Abdallah; Sabine Seeler; Anne Bregnhøj; Gautam Ghatnekar; Lasse S. Kristensen; Lars Iversen; Claus Johansen
• Keywords: keratinocytes; 0301 basic medicine; HSP90 Heat-Shock Proteins/antagonists & inhibitors; mouse model; Immunology; small molecule; Anti-Inflammatory Agents; Antineoplastic Agents; Dermatitis; Dermatitis/drug therapy; Anti-Inflammatory Agents/therapeutic use; Skin Diseases; HSP90 (heat shock protein 90); Mice; 03 medical and health sciences; Psoriasis/drug therapy; Animals; Humans; Psoriasis; HSP90 Heat-Shock Proteins; 12-O-Tetradecanoylphorbol-13-acetate; Inflammation; Interleukin-6; Inflammation/metabolism; Antineoplastic Agents/therapeutic use; RC581-607; novel therapeutic strategy; 3. Good health; inflammation; Quality of Life; Skin Diseases/drug therapy; Immunologic diseases. Allergy

IntroductionChronic inflammatory skin diseases may have a profound negative impact on the quality of life. Current treatment options may be inadequate, offering an unsatisfactory response or side effects. Therefore, ongoing efforts exist to identify novel effective and safe treatments. Heat shock protein (HSP) 90 is a chaperone that promotes the activity of a wide range of client proteins including key proinflammatory molecules involved in aberrant inflammation. Recently, a proof-of-concept clinical trial of 13 patients suggested that RGRN-305 (an HSP90 inhibitor) may be an oral treatment for psoriasis. However, HSP90 inhibition may be a novel therapeutic approach extending beyond psoriasis to include multiple immune-mediated inflammatory skin diseases.MethodsThis study aimed to investigate (i) the anti-inflammatory effects and mechanisms of HSP90 inhibition and (ii) the feasibility of topical RGRN-305 administration (new route of administration) in models of inflammation elicited by 12-O-tetradecanoylphorbol-13-acetate (TPA) in primary human keratinocytes and mice (irritative dermatitis murine model).Results/DiscussionIn primary human keratinocytes stimulated with TPA, a Nanostring® nCounter gene expression assay demonstrated that HSP90 inhibition with RGRN-305 suppressed many proinflammatory genes. Furthermore, when measured by quantitative real-time polymerase chain reaction (RT-qPCR), RGRN-305 significantly reduced the gene expression of TNF, IL1B, IL6 and CXCL8. We next demonstrated that topical RGRN-305 application significantly ameliorated TPA-induced skin inflammation in mice. The increase in ear thickness (a marker of inflammation) was significantly reduced (up to 89% inhibition). In accordance, RT-qPCR of the ear tissue demonstrated that RGRN-305 robustly reduced the gene expression of proinflammatory markers (Tnf, Il1b, Il6, Il17A and Defb4). Moreover, RNA sequencing revealed that RGRN-305 mitigated TPA-induced alterations in gene expression and suppressed genes implicated in inflammation. Lastly, we discovered that the anti-inflammatory effects were mediated, at least partly, by suppressing the activity of NF-κB, ERK1/2, p38 MAPK and c-Jun signaling pathways, which are consistent with previous findings in other experimental models beyond skin inflammation. In summary, HSP90 inhibition robustly suppressed TPA-induced inflammation by targeting key proinflammatory cytokines and signaling pathways. Our findings suggest that HSP90 inhibition may be a novel mechanism of action for treating immune-mediated skin disease beyond psoriasis, and it may be a topical treatment option.