IL6 blockade: an effective approach in steroid-refractory immune checkpoint inhibitor-related neuropathies requiring ICI rechallenge—case report

Background: Colorectal cancer (CRC) is the 4th most common cancer in the US, and the liver is the most common site of metastatic disease. Immune checkpoint inhibitor therapy has not been successful in achieving a clinical response in most patients with CRC liver metastases (CRCLM). The liver is known to induce tolerance to foreign antigens as a result of immunosuppressive cytokines including IL-10. We hypothesized that blockade of IL-10 signaling in CRCLM would potentiate tumor infiltrating lymphocyte (TIL)-mediated tumor cell death. Methods: We performed single-cell RNA sequencing (scRNAseq) of CRCLM using the 10x platform to evaluate for expression of IL-10 or IL-10 receptor (IL-10R) RNA within the tumor (n=8). To confirm if the IL-10R protein was present within the tumor microenvironment (TME), we also performed immunohistochemistry (IHC) (n=3). In order to study the functional effects of IL-10 blockade, we utilized a tumor slice culture (TSC) model, which allows for the study of cancers with their intact TME including immune cells. For TSCs, cores (6 mm diameter) were taken from freshly resected sterile human CRCLM and cut to 250 µm thick slices using a vibratome (n=3). Duplicate slices were treated with either IgG control or anti-IL-10 monoclonal antibodies and cultured for up to 6 days. To evaluate for histological evidence of necrosis and cell apoptosis within the tumor slice, we stained slides with either hematoxylin and eosin (H&E) or cleaved-Caspase-3 (CC3). To gain insight into the activation state of TIL after treatment, we measured levels of cytokines within the culture supernatants. Results: We found by scRNAseq that that IL-10 was expressed by a subset of tumor-associated macrophages, and IL-10R was expressed by both CD4+ and CD8+ T cells as well as macrophages. We confirmed that IL-10R protein was present within the CRCLM TME by IHC, and IL-10R expression was distributed throughout the stroma in non-tumor cells. In TSC treated with anti-IL-10 antibody, CC3+ cells were found to be 82.8% of total cells, compared to 36.1% of control (p = 1 x 10-6) at day 6. These findings were consistent across all human tumor samples treated with IL-10 blockade versus control at all time points examined. Furthermore, IL-10 blockade led to histologic evidence of generalized necrosis compared to an intact TME seen in the control group. Analysis of cytokines released into the media confirmed that IL-10 was present in controls, but absent in slices blocked with anti-IL-10 antibody. We also found increased levels of granzyme B, IL-2, GM-CSF, and IL-18, as well as a reduction in the immune checkpoint receptor TIM3, after one day of IL-10 blockade in culture. Conclusion: Treatment of human CRCLM TSCs with anti-IL-10 antibody leads to a marked increase in immune-mediated cell death within the tumor. Our data suggest that IL-10 serves as a critical regulator of anti-tumor immunity in the CRCLM TME and may serve as an important immunotherapeutic target. Citation Format: Kevin M. Sullivan, Xiuyun Jiang, Yongwoo David Seo, Heidi L. Kenerson, Xiaowei Yan, Chris Lausted, Changting Meng, Neda Jabbari, Kevin P. Labadie, Sara K. Daniel, Qiang Tian, Teresa S. Kim, Raymond S. Yeung, Venu G. Pillarisetty. IL-10 blockade reactivates antitumor immunity in human colorectal cancer liver metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4489.

Lung cancer is the leading cause of cancer death and the second most common cancer diagnosed worldwide in both males and females. K-ras gene mutations are among the most common mutations encountered in non-small cell lung cancer (NSCLC), encompassing up to 30% of them, and are unfortunately associated with chemoresistance and poor prognosis. Recently, immunotherapy treatment of several cancers including NSCLC, particularly by targeting the co-inhibitory molecules program cell death protein 1 (PD-1) and program death-ligand 1 (PD-L1), have shown promising results given either in combination with standard chemotherapy or as neoadjuvant therapy. We have previously shown an important role for IL-6 in K-ras mutant lung tumorigenesis by reprogramming myeloid contexture in the lung tumor microenvironment (TME) and promoting tumor cell proliferation and angiogenesis through the activation of STAT3 pathway. Accordingly, we hypothesized that targeting both immunosuppressive TME and IL-6 driven pro-tumor immune response, through the use of immune checkpoint blockade (ICB), and inhibition of IL-6 would give us a synergistic/additive response and increases the tumor inhibitory effect of ICB. Briefly, starting at the age of 6 weeks, 4 cohorts of K-ras mutant mice (LSL-K-rasG12D/ CCSPCre) (CC-LR) were injected intraperitoneally with vehicle alone (PBS1x), anti-PD-1 (CD279) antibody alone (Clone: 29F.1A12- Bioxcell - BE0273-CUST) (200µg), anti-IL-6 antibody alone (Clone: MP5-20F3 - Bioxcell - BE0046-CUST) (20 mg/kg), or a combination of both. After 8 weeks of treatment, tumor burden and inflammation were studied and quantified. We found a significant (2-fold) decrease in tumor burden in the anti-PD1 alone treated group which was further reduced in the combined anti-PD-1 and anti-IL-6 treated group compared to age and sex matched vehicle treated group. There was also a significant difference in the lung macrophage infiltration in combined anti-PD1/anti-IL-6 treated group similar to what we previously observed in anti-IL-6 alone treated group. Our results suggest that immunomodulatory approaches by targeting cytokine networks like IL-6 blockade could be used to increase the efficacy of immunotherapy, i.e. PD-1 blockade, as a preventive and/or therapeutic strategy for K-ras mutant lung cancer. Citation Format: Marco A. Ramos-Castaneda, Neha Daga, Stephen Peng, Shanshan Deng, Walter Velazco, Mauricio Da Silva Caetano, Seyed Moghaddam. Effect of combined PD-1 and IL-6 blockade on K-ras mutant lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2712.

Tocilizumab for Antibody-Mediated Rejection in the Setting of Cardiac Allograft Vasculopathy

BackgroundMDA-7/IL-24 cytokine has shown potent antitumor properties in various types of cancer without exerting any significant toxicity on healthy cells. It has also been proved to encompass pro-immune Th1 cytokine-like behavior. Several E7 DNA vaccines have developed against human papillomavirus (HPV)-related cervical cancer. However, the restricted immunogenicity has limited their clinical applications individually. To address this deficiency, we investigated whether combining the E7 DNA vaccine with MDA-7/IL-24 as an adjuvant would elicit efficient antitumor responses in tumor-bearing mouse models. Next, we evaluated how suppression of immunosuppressive IL-10 cytokine would enhance the outcome of our candidate adjuvant vaccine.MethodsFor this purpose, tumor-bearing mice received either E7 DNA vaccine, MDA-7/IL-24 cytokine or combination of E7 vaccine with MDA-7/IL-24 adjuvant one week after tumor challenge and boosted two times with one-week interval. IL-10 blockade was performed by injection of anti-IL-10 mAb before each immunization. One week after the last immunization, mice were sacrificed and the treatment efficacy was evaluated through immunological and immunohistochemical analysis. Moreover, the condition of tumors was monitored every two days for six weeks intervals from week 2 on, and the tumor volume was measured and compared within different groups.ResultsA highly significant synergistic relationship was observed between the E7 DNA vaccine and the MDA-7/IL-24 cytokine against HPV-16+ cervical cancer models. An increase in proliferation of lymphocytes, cytotoxicity of CD8+ T cells, the level of Th1 cytokines (IFN-γ, TNF-α) and IL-4, the level of apoptotic markers (TRAIL and caspase-9), and a decrease in the level of immunosuppressive IL-10 cytokine, together with the control of tumor growth and the induction of tumor regression, all prove the efficacy of adjuvant E7&IL-24 vaccine when compared to their individual administration. Surprisingly, vaccination with the DNA E7&IL-24 significantly reduced the population of Regulatory T cells (Treg) in the spleen of immunized mice compared to sole administration and control groups. Moreover, IL-10 blockade enhanced the effect of the co-administration by eliciting higher levels of IFN-γ and caspase-9, reducing Il-10 secretion and provoking the regression of tumor size.ConclusionThe synergy between the E7 DNA vaccine and MDA-7/IL-24 suggests that DNA vaccines’ low immunogenicity can be effectively addressed by coupling them with an immunoregulatory agent. Moreover, IL-10 blockade can be considered a complementary treatment to improve the outcome of conventional or novel cancer therapies.

In the current burgeoning biologic era of autoimmunity therapeutics, an expanding array of specific molecular targets are being used to dismantle the components of the dysregulated immune system felt to be responsible for tissue injury. We are learning that what works well for one disease may not

IntroductionAutoinflammatory fever syndromes are rare and present significant diagnostic and treatment challenges. We present a case which illustrates some key concepts regarding the diagnosis and treatment of a patient with an autoinflammatory disorder, and also touches on the management of active autoinflammation in pregnancy.Case descriptionA 29-year-old woman with a recently identified autoinflammatory disorder was referred to the local rheumatology service in September 2018. She reported having been symptomatic of fever, rashes and arthralgia since the age of 10, and TNF receptor 1 associated periodic syndrome (TRAPS) was suspected on the basis of an N71 deletion on axon 2 of the TNFRS1A gene when tested by a national reference centre 9 months previously.At the time of presentation she was 25 weeks into her sixth pregnancy, the first with a new partner. She reported significant pregnancy morbidity having given birth to a daughter with multiple congenital abnormalities who unfortunately died two days after birth. She suffered 4 subsequent miscarriages at 5-7 weeks gestation with the same partner and underwent extensive genetic testing. At her initial review in the obstetric clinic, she was already receiving low-molecular-weight heparin with aspirin. Colchicine 1 mg tds, did not confer significant symptomatic benefit.One month later, with careful counselling about pregnancy exposure to this biological treatment, IL-1 receptor antagonist (anakinra) therapy was instituted but discontinued after three weeks for generalised rash, as well as lack of efficacy manifesting in raised inflammatory markers. Infection was excluded during a subsequent hospital admission, and prednisolone treatment resulted in significant improvement in clinical course and acute phase response. The patient gave birth to a healthy infant at 37 weeks’ gestation.In the postpartum period, a recurrence of symptoms was observed. IL-6 receptor antagonist (tocilizumab) treatment was commenced at 12 weeks postpartum but discontinued owing to reports of sore throat, cough, headache and fever on the day of the injection. The patient had also discontinued prednisolone on the day of the injection and tocilizumab was rechallenged with good symptomatic response, normalisation of inflammatory markers, and successful reduction in prednisolone dose.DiscussionThis case highlights some interesting points with relation to the treatment of autoinflammatory disorders refractory to IL-1 pathway blockade, and also, of the management of flares of autoinflammation during pregnancy.Firstly, this lady failed to respond to IL-1 receptor antagonist anakinra which has been associated with efficacy in several reports for individuals with TRAPS, and has supplanted TNF blockade with agents such as etanercept for this condition. Only a handful of case reports describe successful IL-6 inhibitor administration for this condition, and this merits further study. This patient’s flare of autoinflammation was treated in the post-partum period, but the demographic characteristics of the autoinflammatory diseases are such that people of child-bearing age may be required to receive treatment in order to prevent pregnancy morbidity as a result of uncontrolled inflammation. IL-6 blockade has not to date been associated with adverse pregnancy outcomes, although this data requires extension and validation. 2 cases of renal agenesis have been reported in children born to mothers with anakinra, but the very small numbers of exposed parents warrants further examination of this observation and the careful study of ongoing pharmacovigilance data to explore this observation.Secondly, success with IL-6 inhibition has been reported in a series of patients with familial Mediterranean fever, the most common autoinflammatory disorder, but this treatment does not feature in the most recent European guidelines for this condition. IL-6 inhibition is well-established in the treatment of adult-onset Still’s disease (AoSD) however, and this lady’s presentation shares important features with that condition. The existence of TNF-receptor 1 mutations has been reported in patients with AoSD, and it may be that this patient’s presentation is more akin to adult-onset Still’s disease and that her TNF receptor mutation is an incidental finding.Key learning pointsThis report adds to the handful cases in which IL-6 blockade has been used successfully to treat the autoinflammatory manifestations of suspected TNF receptor 1 associated periodic syndrome (TRAPS) after the failure of IL-1 receptor blockade with anakinra.Careful history and examination are required to differentiate between potentially overlapping symptoms of drug reaction, autoinflammation, and infection in patients with systemic autoinflammatory disorders (SAIDs). Once a history consistent with autoinflammatory flare was established, cautiously rechallenging with an IL-6 pathway inhibitor ensured this effective treatment was not discounted due to concerns over a drug reaction.With a degree of commonality between the more common conditions in the SAID family, a detailed examination of the clinical phenotype is essential to the interpretation of genetic tests used for the diagnosis of TRAPS and related disorders.Uncontrolled inflammatory disease is associated with pregnancy morbidity and very limited information is available about the short and long-term safety of treatments for autoinflammatory diseases in general. Current guidelines do not recommend the use of IL-6 inhibitor therapy in pregnancy.Conflict of interestThe authors declare no conflicts of interest.

The spectrum of periodic fever syndromes (PFS)/autoinflammation diseases is continuously expanding. This review provides an overview of the primary research and an update on the main clinical developments in these disorders published in the past 12-18 months. IL-1β is pivotal to the pathogenesis of most of the PFS. In familial Mediterranean fever (FMF) MEFV mutations lead to gain of pyrin function, resulting in inappropriate IL-1β release that is dependent on ASC but not the NLRP3 inflammasome. Anti-IL-1 therapy is effective in tumour necrosis factor receptor-associated periodic syndrome (TRAPS), whilst both spontaneous and pathogen-associated molecular patterns (PAMPs) induced IL-1β release have been demonstrated in NLRP12-associated periodic syndrome (NAPS12). Somatic NLRP3/CIAS1 mosaicism is a significant cause of cryopyrin-associated periodic syndromes (CAPS). Close connections have also been established between metabolic and inflammatory pathways. In TRAPS increased reactive oxygen species (ROS) of mitochondrial origin leads to production of pro-inflammatory cytokines, whilst NLRP3 inflammasome activation in type 2 diabetes (T2D) is induced by oligomers of islet amyloid polypeptides (IAPP). Caspase 1 activation and IL-1β release is central to the pathogenesis of many autoinflammatory syndromes. This is supported by the effectiveness of anti-IL-1 biologics in treatment of these disorders.

IL-1 blockade in Schnitzler syndrome: Ex vivo findings correlate with clinical remission

Introduction/Background Dupilumab is a human monoclonal antibody of the IgG4 subclass and binds to the IL-4R alpha subunit, causing downstream inhibition of IL-4 and IL-13 signaling and thus downregulating the T-helper-2 (Th2) cytokine response.1,2,3 Approved by the Food and Drug Administration in 2017, dupilumab is FDA-approved to treat atopic dermatitis (moderate-to-severe), asthma (moderate-to-severe), chronic rhinosinusitis with nasal polyposis, and eosinophilic ­esophagitis. Vitiligo is a common autoimmune depigmenting skin disorder that is prevalent in 1.38% of US adults, and 2.16% in adolescents in the US, with a range of 0.4-2% in most populations.4,5,6 It occurs when the immune system of the body attacks melanocytes, skin cells, that produce melanin, and has been linked to the Koebner phenomenon which occurs when trauma induces lesions, with events including scratching as would be noted in AD.7 AD has also been linked to Vitiligo, especially in children under the age of 12 years.8,9,10 There are many available therapies for vitiligo, topical, systemic, phototherapy, and surgical types being most commonly used.11,12,13 Objectives To address specific features of AA/ Vitiligo/ AD overlap that would support benefit of dupilumab prescribing. Methods An IRB-exempt review was conducted of patient charts for individuals who received dupilumab who had alopecia areata or vitiligo associated with AD. Results Six patients with AD and alopecia universalis (AU), and one patient with AD, vitiligo, and alopecia areata (AA) were identified for review. The patients treated for AU included a 5 year-old Asian female, a 12 year-old white female, a 14 year-old African American male, A 15 year-old Hispanic female, a 26 year-old African American female, and a 78 year-old Hispanic female. The five year-old regrew hair for 4 months but had rapid loss when forced to discontinue due to insurance reasons. The 12 year-old female who had AU for 8 years and had no regrowth. The 15 year-old, 26 year-old, and 78 year-old had rapid regrowth of hair in addition to AD improvement, however, the 15 year-old required addition of an oral JAK inhibitor to retain hair growth. The vitiligo/AA patient is a 61 year-old female patient who had 50% BSA confetti lesion vitiligo affecting the chest, back, abdomen, arms, and legs. She had rapid disease stabilization (2 months) and at 1 year had 90% facial and 70% extremity repigmentation with dupilumab and topical 1.5% ruxolitinib. The same patient had 40% scalp hair loss which fully resolved upon repigmentation of the scalp. Response was noted rapidly but plateaued at 12 months. Conclusions The overlap of AD with AA and Vitiligo points to a shared pathogenesis of the conditions. One of the likely reasons for this is the Koebner phenomenon, which has been poorly characterized. We hypothesize that the Koebner phenomenon is triggered through an IL-4/ IL-13 mechanism, i.e. for specific individuals IL-4 and IL-13 hyper-reactivity can be a Koebner-based trigger. Additionally, AA overlaps with AD is poorly characterized. However, the linkage has been recognized in recent AAD guidelines addressing AD comorbidities.14 We hypothesize that IL-4/ IL-13 hyper-reactivity in the skin can act similarly in AU. There is in AA an overlap of TH1-CXCL9/10 expression and interferon gamma overexpression in addition to TH2- including IL-13 overexpression.15 Therefore, the blockade of IL-4/ IL-13 may be effective through multiple mechanisms of activity. There is already notable Phase 2a data supporting dupilumab usage in AA.16 Our experience demonstrates sustained hair growth in half of patients treated. AA/ Vitiligo overlap appears to respond well to dupilumab. This is supported by a recent case report demonstrating benefit of dupilumab in programmed cell death inhibitor-1 induced vitiligo with associated refractory pruritus.17 Liu et al have observed in vitro that rising IL-4 levels were linked to increased vitiligo risk.18 On the other hand, some reports of vitiligo after initiating dupilumab therapy do exist, with some new-onset and worsening described. These cases are limited but bear consideration. In our patient, disease stabilization was noted in a generalized confetti-vitiligo patient, but a topical JAK inhibitor was used adjunctively.19,20,21,22,23 Given the need for long-term maintenance, the safety of dupilumab is an attractive feature. Therefore, further exploration of dupilumab therapy for vitiligo/AA/AD, vitiligo/AD/Koebner+, and AU/ AD is needed. This is particularly important to address in patients under the age of 12 years who currently have no approved systemic medications for vitiligo and AU.24 When AU, vitiligo, and combinations of the two are comorbid with AD, there is an expectation of circulating IL-4/13 elevations and localized IL-4/13 elevation that support a potential role for dupilumab therapy in these conditions. Given that AD is associated with vitiligo of childhood, and severe alopecia areata is linked to AD, systemic therapy would be necessary in these individuals.

Background: Dupilumab is used for treatment of atopic dermatitis through blockade of IL-4 and IL-13 signaling of the Th2 pathway. Recent case reports have described alopecia, psoriasis, persistent facial dermatitis, and recall dermatitis at patch test sites after the initiation of dupilumab therapy.Case report: We describe the case of a 67-year-old female with atopic dermatitis who developed recurrent episodic flares of rosacea temporally associated with dupilumab injections that resolved after dupilumab discontinuation.Conclusion: The cause of rosacea-like reaction associated with dupilumab treatment is unknown. Th2 pathway inhibition by dupilumab may promote Demodex proliferation and increased IL-17-mediated inflammation implicated in the pathophysiology of rosacea.Abbreviations: atopic dermatitis: AD; interleukin: IL; persistent facial dermatitis: PFD; T-helper cell type 1: Th1; T-helper cell type 2: Th2; T-helper cell type 17: Th17; tumor necrosis factor-∝: TNF-∝

BackgroundMevalonate kinase deficiency (MKD) is a rare autoinflammatory condition caused by biallelic loss-of-function (LOF) mutations in mevalonate kinase (MVK) gene encoding the enzyme mevalonate kinase. Patients with MKD display a variety of non-specific clinical manifestations, which can lead to diagnostic delay. We report the case of a child presenting with vasculitis that was found by genetic testing to be caused by MKD, and now add this autoinflammatory disease to the ever-expanding list of causes of monogenic vasculitides.Case presentationA 2-year-old male presented with an acute 7-day history of high-grade fever, abdominal pain, diarrhoea, rectal bleeding and extensive purpuric and necrotic lesions, predominantly affecting the lower limbs. He had been suffering from recurrent episodes of fever from early in infancy, associated with maculopapular/petechial rashes triggered by intercurrent infection, and after vaccines. Extensive infection screen was negative. Skin biopsy revealed small vessel vasculitis. Visceral digital subtraction arteriography was normal. With a diagnosis of severe idiopathic cutaneous vasculitis, he was treated with corticosteroids and mycophenolate mofetil. Despite that his acute phase reactants remained elevated, fever persisted and the vasculitic lesions progressed. Next-generation sequencing revealed compound heterozygous mutation in MVK c.928G > A (p.V310M) and c.1129G > A (p.V377I) while reduced mevalonate enzyme activity was confirmed suggesting a diagnosis of MKD as a cause of the severe vasculitis. Prompt targeted treatment with IL-1 blockade was initiated preventing escalation to more toxic vasculitis therapies and reducing unnecessary exposure to cytotoxic treatment.ConclusionsOur report highlights the broad clinical phenotype of MKD that includes severe cutaneous vasculitis and emphasizes the need to consider early genetic screening for young children presenting with vasculitis to exclude a monogenic vasculitis which may be amenable to targeted treatment.

Muckle-Wells syndrome (MWS), a rare autoinflammatory disorder within the cryopyrin-associated periodic syndrome (CAPS) spectrum, is primarily characterized by recurrent fevers, urticarial rash, sensorineural hearing loss, and risk of amyloidosis. Although systemic manifestations are well-documented, gastrointestinal (GI) symptoms remain underrecognized and poorly described. This systematic review explores the prevalence, diagnostic relevance, and treatment response of GI manifestations in MWS. A structured search strategy was employed using major databases, and studies were included if they involved patients with genetically or clinically confirmed MWS and reported GI symptoms such as abdominal pain or oral ulcers. A total of three studies met the inclusion criteria, including two observational cohorts and one case report. Abdominal pain was noted in up to one-third of patients with childhood-onset disease and recurrently in a confirmed case. While IL-1 blockade with anakinra or canakinumab demonstrated overall systemic improvement, GI outcomes were not consistently reported. These findings suggest that gastrointestinal involvement, though infrequently highlighted, may be clinically significant and should be integrated into diagnostic and therapeutic frameworks for MWS.

Refractory Kawasaki disease (KD) is related to a major risk of coronary arteries abnormalities and its treatment is not standardized. In this regard, anakinra (ANA), an interleukin (IL)-1 receptor antagonist, represents an emerging therapeutic option. We report two cases of children, diagnosed with KD, nonresponsive to two doses of intravenous immunoglobulins, successfully treated with ANA, without a prior use of steroids. Patient 2 developed a coronary dilatation, that improved significantly after ANA therapy. Our experience highlights IL-1 blockade effectiveness in reducing KD inflammation and suggests ANA adoption as second-line therapy, with a timesaving and steroid-sparing strategy. Our results, combined with the evidence of the IL-1 key role in KD and coronary arteritis pathogenesis and to the recent clinical evidence reported by the KAWAKINRA trial, encourage an earlier recourse to ANA in patients with refractory KD, in order to fight inflammation, and to treat and prevent the development of coronary artery aneurysms. Further studies are needed to better define the place of IL-1 blockade in KD step-up treatment.

BACKGROUND:Currently, there are no therapies targeting specific pathogenic pathways in myocarditis. IL (interleukin)-1 blockade has shown promise in preclinical studies and case reports. We hypothesized that blockade of IL1RAP (IL-1 receptor accessory protein), a shared subunit of the IL-1, IL-33, and IL-36 receptors, could be more efficient than IL-1 blockade alone.METHODS:We induced coxsackievirus B3 (CVB3)–mediated or experimental autoimmune myocarditis (EAM) in BALB/c mice, followed by treatment with an Fc (fragment crystallizable)-modified mIgG2a mouse anti-mouse IL1RAP monoclonal antibody (mCAN10). Myocarditis severity and immune infiltration were assessed by histology and flow cytometry. Cardiac function was measured by echocardiography. We used spatial transcriptomics (Visium 10× Genomics) to compare the gene expression landscape in the hearts of mCAN10-treated versus control mice.RESULTS:IL1RAP blockade reduced CVB3 and EAM severity. In EAM, the treatment prevented deterioration of cardiac function, measured on day 42 post-disease induction (left ventricular ejection fraction: 56.5% versus 51.0% in isotype controls [P=0.002] and versus 51.4% in mice treated with anti-IL-1β antibodies alone [P=0.003]; n=10–11 mice per group). In the CVB3 model, mCAN10 did not impede viral clearance from the heart and significantly lowered the numbers of CD4+ (cluster of differentiation 4) T cells (P=0.025), inflammatory Ly6C+CCR2+ (lymphocyte antigen 6 complex, locus C/C-C motif chemokine receptor 2) monocytes (P=0.038), neutrophils (P=0.001) and eosinophils (P<0.001) infiltrating the myocardium. The spatial transcriptomic analysis revealed reduced canonical IL-1 signaling and chemokine expression in cardiac immune foci in CVB3-infected mice treated with IL1RAP blockade.CONCLUSIONS:Blocking IL1RAP reduces acute CVB3 myocarditis and EAM severity and preserves cardiac function in EAM. We conclude that IL1RAP blockade is a potential therapeutic strategy in viral and autoimmune myocarditis.

First reported case of safe and efficacious use of tocilizumab for treatment of hyperinflammatory syndrome associated with COVID‐19 in an allogeneic stem cell transplant patient