Specific activation of pro-Infliximab enhances selectivity and safety of rheumatoid arthritis therapy

Lu, Yun-Chi; Chuang, Chih-Hung; Chuang, Kuo-Hsiang; Chen, I-Ju; Huang, Bo-Cheng; Lee, Wen-Han; Wang, Hsin‐Ell; Li, Jia-Je; Cheng, Yi-An; Cheng, Kai-Wen; Wang, Jaw‐Yuan; Hsieh, Yuan-Chin; Lin, Wen-Wei; Cheng, Tian−Lu

doi:10.1371/journal.pbio.3000286

Cited by 23 publications

(26 citation statements)

References 44 publications

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“…Three weeks after intravenous inoculation of 1.5×10ˆ6 p401 PDX tumor cells/mouse, we injected CLL-1 CAR +iC9-IL15 CAR T cells (2.5×10ˆ6/mouse) T cells ( figure 5A ), followed by intravenous injections of anti-TNFα. 24 25 The TNFα levels were significantly reduced in mice treated with the first dose of anti-TNFα ( figure 5B ) but were not well controlled by subsequent injections. This treatment had no adverse effect on T cell expansion or tumor control ( figure 5C ).…”

Section: Resultsmentioning

confidence: 96%

Modulating TNFα activity allows transgenic IL15-Expressing CLL-1 CAR T cells to safely eliminate acute myeloid leukemia

Atilla

McKenna

Tashiro

et al. 2020

J Immunother Cancer

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BackgroundC-type lectin-like molecule 1 (CLL-1) is highly expressed in acute myeloid leukemia (AML) but is absent in primitive hematopoietic progenitors, making it an attractive target for a chimeric antigen receptor (CAR) T-cell therapy. Here, we optimized our CLL-1 CAR for anti-leukemic activity in mouse xenograft models of aggressive AML.MethodsFirst, we optimized the CLL-1 CAR using different spacer, transmembrane and costimulatory sequences. We used a second retroviral vector to coexpress transgenic IL15. We measured the effects of each construct on T cell phenotype and sequential (recursive) co culture assays with tumor cell targets to determine the durability of the anti tumor activity by flow cytometry. We administered CAR T cells to mice engrafted with patient derived xenografts (PDX) and AML cell line and determined anti tumor activity by bioluminescence imaging and weekly bleeding, measured serum cytokines by multiplex analysis. After euthanasia, we examined formalin-fixed/paraffin embedded sections. Unpaired two-tailed Student’s t-tests were used and values of p<0.05 were considered significant. Survival was calculated using Mantel-Cox log-rank test.ResultsIn vitro, CLL-1 CAR T cells with interleukin-15 (IL15) were less terminally differentiated (p<0.0001) and had superior expansion compared with CD28z-CD8 CAR T cells without IL15 (p<0.001). In both AML PDX and AML cell line animal models, CLL-1 CAR T coexpressing transgenic IL15 initially expanded better than CD28z-CD8 CAR T without IL15 (p<0.0001), but produced severe acute toxicity associated with high level production of human tumor necrosis factor α (TNFα), IL15 and IL2. Histopathology showed marked inflammatory changes with tissue damage in lung and liver. This acute toxicity could be managed by two strategies, individually or in combination. The excessive TNF alpha secretion could be blocked with anti-TNF alpha antibody, while excessive T cell expansion could be arrested by activation of an inducible caspase nine safety switch by administration of dimerizing drug. Both strategies successfully prolonged tumor-free survival.ConclusionCombinatorial treatment with a TNFα blocking antibody and subsequent activation of the caspase-9 control switch increased the expansion, survival and antileukemic potency of CLL-1 CAR T-cells expressing transgenic IL15 while avoiding the toxicities associated with excessive cytokine production and long-term accumulation of activated T-cells.

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Section: Resultsmentioning

confidence: 96%

Modulating TNFα activity allows transgenic IL15-Expressing CLL-1 CAR T cells to safely eliminate acute myeloid leukemia

Atilla

McKenna

Tashiro

et al. 2020

J Immunother Cancer

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“…Starting drug treatment in the early stage can effectively prevent the progression of the disease and reduce the rate of disease development. Because a large number of reports on the regenerative function of Treg cells have been published and the ideal treatment strategy is to induce self-tolerance before obvious tissue damage occurs, researchers have designed various strategies ways to increase the number of Treg cells and restore their function(101)(102)(103)(104)(105)(106)(107), by enhancing the function of Treg cells in vivo, including reducing the pro-inflammatory environment and enhancing the response of effector cells to inhibition (108-113) (Table…”

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confidence: 99%

Function and Role of Regulatory T Cells in Rheumatoid Arthritis

et al. 2021

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Rheumatoid arthritis (RA) is a systemic and heterogeneous autoimmune disease with symmetrical polyarthritis as its critical clinical manifestation. The basic cause of autoimmune diseases is the loss of tolerance to self or harmless antigens. The loss or functional deficiency of key immune cells, regulatory T (Treg) cells, has been confirmed in human autoimmune diseases. The pathogenesis of RA is complex, and the dysfunction of Tregs is one of the proposed mechanisms underlying the breakdown of self-tolerance leading to the progression of RA. Treg cells are a vital component of peripheral immune tolerance, and the transcription factor Foxp3 plays a major immunosuppressive role. Clinical treatment for RA mainly utilizes drugs to alleviate the progression of disease and relieve disease activity, and the ideal treatment strategy should be to re-induce self-tolerance before obvious tissue injury. Treg cells are one of the ideal options. This review will introduce the classification, mechanism of action, and characteristics of Treg cells in RA, which provides insights into clinical RA treatment.

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“…[4] These clinical ndings indicated that preventing the interference effect of anti-Id Ab can solve many of the clinical problems of Ab drugs. Our team have demonstrated that the spatial hindrance-based Ab lock can prevent the response of anti-Id Ab to In iximab (anti-I-Id Ab); the binding of the anti-I-Id Ab to pro-In iximab was 108-fold weaker than that to In iximab [19], and the Ab lock maintained the TNFa binding ability of pro-In iximab which was pre-incubated with anti-Id Ab. In the future, we will be able to e ciently transform Abs into optimized pro-Abs to prevent the binding of anti-Id Ab using MSCS.…”

Section: Discussionmentioning

confidence: 99%

“…Further, pro-In iximab not only showed equivalent therapeutic e cacy to In iximab but also maintained immunity against Listeria infection in the RA mouse model, which led to a 71% survival rate compared to 0% of the In iximab treatment group, indicating that pro-Abs can enhance Ab selectivity and reduce side effects. [19] Our previous results showed that by using the concept of "copy and paste", we transformed almost 70% of therapeutic Abs into pro-Abs and achieved over 100-fold blocking ability compared to the parental Abs. Hence, this strategy could be further applied to develop customized pro-Abs.…”

Section: Introductionmentioning

confidence: 99%

Development of a Structure-based Computational Simulation to Optimize the Blocking Efficacy of Pro-antibodies

Huang

Liao

et al. 2020

Preprint

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Background: The on-target toxicity of monoclonal antibodies (Abs) is mainly due to the fact that Abs cannot distinguish target antigens (Ags) expressed in disease regions from those in normal tissues during systemic administration. In order to overcome this issue, we “copied” an autologous Ab hinge as an “Ab lock” and “pasted” it on the binding site of the Ab by connecting a protease substrate and linker in between to generate a pro-Ab, which can be specifically activated in the disease region to enhance Ab selectivity and reduce side effects. Previously, we reported that 70% of pro-Abs can achieve more than 100-fold blocking ability compared to the parental Abs. However, 30% of pro-Abs do not have such efficient blocking ability. This is because the same Ab lock linker cannot be applied to every Ab due to the differences in the complementarity-determining region (CDR) loops. Here we designed a method which uses structure-based computational simulation (MSCS) to optimize the blocking ability of the Ab lock for all Ab drugs. MSCS can precisely adjust the amino acid composition of the linker between the Ab lock and Ab drug with the assistance of molecular simulation.Results: We selected αPD-1, αIL-1β, αCTLA-4 and αTNFα Ab as models and attached the Ab lock with various linkers (L1 to L7) to form pro-Abs by MSCS, respectively. The resulting cover rates of the Ab lock with various linkers compared to the Ab drug were in the range 28.33%-42.33%. The recombinant pro-Abs were generated by MSCS prediction in order to verify the application of molecular simulation for pro-Ab development. The binding kinetics effective concentration (EC-50) for αPD-1 (200-250-fold), αIL-1β (152-186-fold), αCTLA-4 (68-150-fold) and αTNFα Ab (20-123-fold) were presented as the blocking ability of pro-Ab compared to the Ab drug. Further, there was a positive correlation between cover rate and blocking ability of all pro-Ab candidates. Conclusions: The results suggested that MSCS was able to predict the Ab lock linker most suitable for application to αPD-1, αIL-1β, αCTLA-4 and αTNFα Ab to form pro-Abs efficiently. The success of MSCS in optimizing the pro-Ab can aid the development of next-generation pro-Ab drugs to significantly improve Ab-based therapies and thus patients’ quality of life.

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Specific activation of pro-Infliximab enhances selectivity and safety of rheumatoid arthritis therapy

Cited by 23 publications

References 44 publications

Modulating TNFα activity allows transgenic IL15-Expressing CLL-1 CAR T cells to safely eliminate acute myeloid leukemia

Modulating TNFα activity allows transgenic IL15-Expressing CLL-1 CAR T cells to safely eliminate acute myeloid leukemia

Function and Role of Regulatory T Cells in Rheumatoid Arthritis

Development of a Structure-based Computational Simulation to Optimize the Blocking Efficacy of Pro-antibodies

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