Presentation Date

19-10-2021 12:00 AM

Description

Background: Osteoarthritis is a complex multifactorial disorder that is the most frequent cause of pain, loss of function, and disability in adults1. Many modalities of treatment involve symptom alleviation but typically lead to joint replacement in end-stage disease. Consequently, there has been a shift towards the prevention of disease progression of early osteoarthritis. Due to advances in biochemical marker development, pro-inflammatory interleukins, specifically interleukin 1β (IL1β), has come into focus2. IL1β is implicated in many of the pathological advancement of osteoarthritis such as degradation of cartilage, bony malformation, and thickening of synovial tissue3-5. Previously, IL1 receptor antagonist (IL1ra) proved to be effective at preventing cartilage breakdown and osteoarthritis progression. This led to the development of a drug called Anakinra. Despite the promising results in reduced pain, due to its short half-life, Anakinra proved to be no more effective than a placebo for long term treatment 6-12. To address this issue, Flexion Therapeutics developed FX201, a helper-dependent adenovirus (HDAd) vector to carry the coding sequence for IL1ra under the control of an inflammation-responsive promoter. Aims: This project will determine the efficacy of Flexion’s FX201, a gene therapy product for the delivery of human interleukin 1 receptor antagonist, to reduce inflammatory output of osteoarthritic synoviocytes. Additionally, this project will identify target patient populations for clinical trials with FX201. We hypothesize that FX201 will reduce inflammatory output to a greater extent in cells derived from patients with high circulating levels of IL1, TNF and IFN than in cells from patients with low circulating levels of these same inflammatory cytokines. Methods: 12 patients were selected from Louisiana State University Health Sciences Center biorepository and subdivided into high/low pain scores based on knee injury and osteoarthritis outcome scores (KOOS) and pro-/anti-inflammatory profile based on circulating levels of IL1, TNF and IFN. Synoviocytes from previously frozen synovial tissue were extracted, plated in media, and incubated at 37o C 5% CO2. Synoviocytes were then treated with FX201 and measure cytokine release by cells before and after infection. A multiplex analysis containing 11 pro- and anti-inflammatory cytokines, and 6 proteins involved in cartilage turnover was used to measure secreted protein levels before and after FX201 treatment. Quantitative PCR was secondarily used to confirm decreases in cytokine production. Expected Results: Patient’s extracted synoviocytes will quantitatively show a reduction in pro- inflammatory cytokines and proteins involved in cartilage degradation when treated with FX201. Cells from patients with high levels of inflammation will show a greater effect than cells from patients with low levels of inflammation. 1. Nigel Arden, Michael C. Nevitt, Osteoarthritis: Epidemiology, Best Practice & Research Clinical Rheumatology, Volume 20, Issue 1, 2006, Pages 3-25, ISSN 1521-6942 2. S Glyn-Jones, A J R Palmer, R Agricola, A J Price, T L Vincent, H Weinans, A J Carr, Osteoarthritis,The Lancet, Volume 386, Issue 9991, 2015, Pages 376-387, ISSN 0140-6736 3. Raghu H, Mao R, Lindstrom TM, et al. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis. Nature Reviews Rheumatology. 2016;12(10):580. 4. Van Lent P, Blom A, Van Der Kraan P, Holthuysen A, Vitters E, Van Rooijen N, et al. Crucial role of synovial lining macrophages in the promotion of transforming growth factor β–mediated osteophyte formation. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 2004;50(1):103-11. 5. Glasson SS. In vivo osteoarthritis target validation utilizing genetically-modified mice. Current drug targets. 2007;8(2):367-76. 6. Caron JP, Fernandes JC, Martel‐Pelletier J, Tardif G, Mineau F, Geng C, et al. Chondroprotective effect of intraarticular injections of interleukin‐1 receptor antagonist in experimental osteoarthritis. Suppression of collagenase‐1 expression. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 1996;39(9):1535-44. 7. Pelletier JP, Caron JP, Evans C, Robbins PD, Georgescu HI, Jovanovic D, et al. In vivo suppression of early experimental osteoarthritis by interleukin‐1 receptor antagonist using gene therapy. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 1997;40(6):1012-9. 8. Fernandes J, Tardif G, Martel-Pelletier J, Lascau-Coman V, Dupuis M, Moldovan F, et al. In vivo transfer of interleukin-1 receptor antagonist gene in osteoarthritic rabbit knee joints: prevention of osteoarthritis progression. The American journal of pathology. 1999;154(4):1159-69. 9. Zhang X, Mao Z, Yu C. Suppression of early experimental osteoarthritis by gene transfer of interleukin‐1 receptor antagonist and interleukin‐10. Journal of orthopaedic research. 2004;22(4):742-50. 10. Wang H-j, Yu C-l, Kishi H, Motoki K, Mao Z-b, Muraguchi A. Suppression of experimental osteoarthritis by adenovirus-mediated double gene transfer. Chinese medical journal. 2006;119(16):1365-73. 11. Santangelo K, Nuovo G, Bertone A. In vivo reduction or blockade of interleukin-1β in primary osteoarthritis influences expression of mediators implicated in pathogenesis. Osteoarthritis and cartilage. 2012;20(12):1610-8. 12. Frisbie D, Ghivizzani S, Robbins PD, Evans CH, McIlwraith C. Treatment of experimental equine osteoarthritis by in vivo delivery of the equine interleukin-1 receptor antagonist gene. Gene therapy. 2002;9(1):12-20.

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Oct 19th, 12:00 AM

The Development of a Novel Gene Therapy, Flexion’s FX201, for theTreatment of Osteoarthritis

Background: Osteoarthritis is a complex multifactorial disorder that is the most frequent cause of pain, loss of function, and disability in adults1. Many modalities of treatment involve symptom alleviation but typically lead to joint replacement in end-stage disease. Consequently, there has been a shift towards the prevention of disease progression of early osteoarthritis. Due to advances in biochemical marker development, pro-inflammatory interleukins, specifically interleukin 1β (IL1β), has come into focus2. IL1β is implicated in many of the pathological advancement of osteoarthritis such as degradation of cartilage, bony malformation, and thickening of synovial tissue3-5. Previously, IL1 receptor antagonist (IL1ra) proved to be effective at preventing cartilage breakdown and osteoarthritis progression. This led to the development of a drug called Anakinra. Despite the promising results in reduced pain, due to its short half-life, Anakinra proved to be no more effective than a placebo for long term treatment 6-12. To address this issue, Flexion Therapeutics developed FX201, a helper-dependent adenovirus (HDAd) vector to carry the coding sequence for IL1ra under the control of an inflammation-responsive promoter. Aims: This project will determine the efficacy of Flexion’s FX201, a gene therapy product for the delivery of human interleukin 1 receptor antagonist, to reduce inflammatory output of osteoarthritic synoviocytes. Additionally, this project will identify target patient populations for clinical trials with FX201. We hypothesize that FX201 will reduce inflammatory output to a greater extent in cells derived from patients with high circulating levels of IL1, TNF and IFN than in cells from patients with low circulating levels of these same inflammatory cytokines. Methods: 12 patients were selected from Louisiana State University Health Sciences Center biorepository and subdivided into high/low pain scores based on knee injury and osteoarthritis outcome scores (KOOS) and pro-/anti-inflammatory profile based on circulating levels of IL1, TNF and IFN. Synoviocytes from previously frozen synovial tissue were extracted, plated in media, and incubated at 37o C 5% CO2. Synoviocytes were then treated with FX201 and measure cytokine release by cells before and after infection. A multiplex analysis containing 11 pro- and anti-inflammatory cytokines, and 6 proteins involved in cartilage turnover was used to measure secreted protein levels before and after FX201 treatment. Quantitative PCR was secondarily used to confirm decreases in cytokine production. Expected Results: Patient’s extracted synoviocytes will quantitatively show a reduction in pro- inflammatory cytokines and proteins involved in cartilage degradation when treated with FX201. Cells from patients with high levels of inflammation will show a greater effect than cells from patients with low levels of inflammation. 1. Nigel Arden, Michael C. Nevitt, Osteoarthritis: Epidemiology, Best Practice & Research Clinical Rheumatology, Volume 20, Issue 1, 2006, Pages 3-25, ISSN 1521-6942 2. S Glyn-Jones, A J R Palmer, R Agricola, A J Price, T L Vincent, H Weinans, A J Carr, Osteoarthritis,The Lancet, Volume 386, Issue 9991, 2015, Pages 376-387, ISSN 0140-6736 3. Raghu H, Mao R, Lindstrom TM, et al. Low-grade inflammation as a key mediator of the pathogenesis of osteoarthritis. Nature Reviews Rheumatology. 2016;12(10):580. 4. Van Lent P, Blom A, Van Der Kraan P, Holthuysen A, Vitters E, Van Rooijen N, et al. Crucial role of synovial lining macrophages in the promotion of transforming growth factor β–mediated osteophyte formation. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 2004;50(1):103-11. 5. Glasson SS. In vivo osteoarthritis target validation utilizing genetically-modified mice. Current drug targets. 2007;8(2):367-76. 6. Caron JP, Fernandes JC, Martel‐Pelletier J, Tardif G, Mineau F, Geng C, et al. Chondroprotective effect of intraarticular injections of interleukin‐1 receptor antagonist in experimental osteoarthritis. Suppression of collagenase‐1 expression. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 1996;39(9):1535-44. 7. Pelletier JP, Caron JP, Evans C, Robbins PD, Georgescu HI, Jovanovic D, et al. In vivo suppression of early experimental osteoarthritis by interleukin‐1 receptor antagonist using gene therapy. Arthritis & Rheumatism: Official Journal of the American College of Rheumatology. 1997;40(6):1012-9. 8. Fernandes J, Tardif G, Martel-Pelletier J, Lascau-Coman V, Dupuis M, Moldovan F, et al. In vivo transfer of interleukin-1 receptor antagonist gene in osteoarthritic rabbit knee joints: prevention of osteoarthritis progression. The American journal of pathology. 1999;154(4):1159-69. 9. Zhang X, Mao Z, Yu C. Suppression of early experimental osteoarthritis by gene transfer of interleukin‐1 receptor antagonist and interleukin‐10. Journal of orthopaedic research. 2004;22(4):742-50. 10. Wang H-j, Yu C-l, Kishi H, Motoki K, Mao Z-b, Muraguchi A. Suppression of experimental osteoarthritis by adenovirus-mediated double gene transfer. Chinese medical journal. 2006;119(16):1365-73. 11. Santangelo K, Nuovo G, Bertone A. In vivo reduction or blockade of interleukin-1β in primary osteoarthritis influences expression of mediators implicated in pathogenesis. Osteoarthritis and cartilage. 2012;20(12):1610-8. 12. Frisbie D, Ghivizzani S, Robbins PD, Evans CH, McIlwraith C. Treatment of experimental equine osteoarthritis by in vivo delivery of the equine interleukin-1 receptor antagonist gene. Gene therapy. 2002;9(1):12-20.