Protagen (Germany)

Health Outcomes with Padua Gene; Evaluation in Hemophilia B (HOPE-B) is the first phase 3 trial of adeno-associated virus (AAV) serotype 5 vector-based gene therapy for hemophilia B to have enrolled participants with neutralizing antibodies (NAbs) to the viral vector. Evaluate the efficacy and safety of etranacogene dezaparvovec in a post hoc subgroup of participants with preexisting AAV5 NAbs 4 years posttherapy. After a ≥6-month lead-in period on continuous prophylaxis, people with moderately severe/severe hemophilia B (factor [F]IX ≤ 2%) received a single infusion of etranacogene dezaparvovec. AAV5 NAb status prior to infusion was determined. FIX activity, annualized bleeding rates (ABRs), and safety were evaluated in participants with preexisting AAV5 NAbs. A total of 21/54 participants had detectable preexisting AAV5 NAbs (titer: 8.5-678, n = 20; titer: 3212, n = 1). Two participants did not respond to treatment (high titer: 3212, n = 1; received partial dose, n = 1). The mean FIX activity was 36 IU/dL (SD, 18) at 1 year (n = 18) and 34 IU/dL (SD, 16) at 4 years (n = 15) posttreatment. Unadjusted ABRs for all bleeds (treated and untreated) decreased by 74.4% from 4.64 during lead-in to 1.18 during months 7 to 48 (n = 21). ABRs for spontaneous and joint bleeds decreased by 79% and 82%, respectively. The mean FIX consumption decreased from 245,476 IU/y (SD, 144,497) during lead-in to 23,975 IU/y (SD, 48.928) during years 1 to 4 (P < .0001). The most frequent treatment-related adverse events were infusion-related reactions (23.8%) and transient alanine aminotransferase elevations (14.3%). No late hepatotoxicity was observed. Etranacogene dezaparvovec demonstrated long-term efficacy and safety in individuals with preexisting AAV5 NAb titers ≤ 678, expanding eligibility for gene therapy to a broader hemophilia B population.

BackgroundInterstitial lung disease (ILD) is one of the major causes of morbidity and mortality in patients with idiopathic inflammatory myopathies (IIM). [1] Within IIM, ILD is frequently found in patients with anti-synthetase syndrome (ASyS). Current treatment is based on a combination of high doses of glucocorticoids and other immunosuppressors such as cyclophosphamide, with progression of ILD despite the treatment. One anti-fibrotic drug, nintedanib, has recently been approved for patients with progressive fibrotic ILD.ObjectivesTo explore if within patients with ASyS there is a group with worsening ILD that fits the definition of progressive pulmonary fibrosis (PPF)[2] and thus would be eligible for anti-fibrotic treatment.[3]MethodsA single center based retrospective cohort of patients diagnosed with ASyS, ILD involvement, and positivity for specific autoantibodies anti-Jo1, anti-PL7, anti-PL12, anti-EJ and/or anti-OJ (ASyS-AB) was identified using the Swedish Myositis network register. We selected the latest computed tomography (CT) thoracic scans, pulmonary function tests (PFT) and the development of respiratory symptoms according to clinical records. Patients were evaluated based on the INBUILD study criteria[3] and the international thoracic and respiratory clinical practice guideline (CPG)[2] for the presence of PPF. An experienced thoracic radiologist reviewed the CT scans of those who met the criteria to confirm PPF.ResultsWe identified 147 individuals positive for ASyS-AB; 66 of them were not included due to either a follow-up period of less than one year, lack of data in the national registry or absence of ILD. Finally, a cohort of 81 patients was included, 67.9% women, with a median (Q1,Q3) age at disease onset of 58 (48,67) years (Table 1). The median time from disease onset until assessment for PPF criteria was of 6 years.The most prevalent CT finding amongst all groups was fibrotic changes (n=28) neither fitting the usual interstitial pneumonia (UIP) (n=9) nor the fibrotic non-specific interstitial pneumonia (NSIP) (n=6) criteria; followed by inflammatory NSIP (n= 18). 14.8 percent or 19.8 percent (INBUILD and CPG, retrospectively) of the individuals with any type of lung fibrotic pattern on CT met the definition of PPF.Table 1.Baseline characteristics of patients with anti-synthetase syndrome (ASyS) in the cohortTotalBy antibodyPL7,n=8PL12,n=8Jo1,n=58EJ,n=4OJ,n=3Women, n (%)55 (67.9)6 (75)5 (62.5)40 (69)3 (75)1 (33.3)Median age at disease onset, years (Q1,Q3)58 (48,67)61 (35,68)61 (48,67)56 (47,65)72 (46,76)61 (55,74)Time since disease onset at evaluation, years (Q1,Q3)6 (3,11)10 (3,22)4 (2,5)7 (3,13)2 (2,15)4 (1,7)Patients meeting criteria for progressive fibrosis, n (%)INBUILD criteria12 (14.8)2 (25)1 (12.5)8 (13.8)0 (0)1 (33.3)ATS/ERS/JRS Clinical practice guidelines14 (17.3)3 (37.5)1 (12.5)10 (17.2)0 (0)0 (0)Time since disease onset at evaluation, years (Q1,Q3)10 (5,21)-----ConclusionIn this cross-sectional study, 15% of patients with ASyS fulfilled the definition of progressive fibrosis and thus could be eligible for anti-fibrotic treatment. To our knowledge, this is the first cohort to demonstrate a possible real-life target of anti-fibrotics in IIM-ILD.

ABSTRACTBackgroundImproved knowledge regarding the prevalence and clinical significance of the broad spectrum of autoantibodies triggered by SARS-CoV2 infection can clarify the underlying pathobiology, enhance approaches to evaluating heterogeneity of COVID-19 clinical manifestations, and potentially guide options for targeting immunosuppressive therapy as the need for more effective interventions continues to evolve. In this study, we sought to determine the prevalence of autoimmune antibodies in diverse cohort of SARS-CoV-2 positive healthcare workers and measure the extent to which factors associated with triggered autoimmunity are activated even following mild and asymptomatic infection.MethodsAntigen microarrays were used to profile reactivity of IgG autoantibodies against 91 proteins and cytokines based on autoantibody profiling studies in autoimmune diseases.ResultsIn this discovery screening study, we found that 90% of the IgG positive individuals demonstrated reactivity to at least one autoantibody. When compared to results of the same assays conducted on samples from pre-COVID-19 controls, our primary cohort of individuals with SARS-CoV-2 IgG antibody positivity had significantly elevated IgG against twelve additional proteins including CHD3, CTLA4, HARS, IFNA4, INS, MIF, MX1, RNF41, S100A9, SRP19, TROVE2, and VEGFA. These findings confirmed that all severity levels of SARS-CoV-2 infection, even asymptomatic infections, trigger a robust and diverse autoimmune response; our results also highlight the utility of multiparametric autoantibody detection in this setting.InterpretationTaken together, our findings underscore the serological diversity underlying the clinical heterogeneity of COVID-19 infection and its sequelae, including the long-Covid phenotypes.FundingThis work was supported in part by Cedars-Sinai Medical Center (JEE; SC), the Erika J Glazer Family Foundation (JEE; JEVE; SC), CSMC Precision Health Grant (JFB), the F. Widjaja Family Foundation (JGB, GYM, DM), the Helmsley Charitable Trust (JGB, GYM, DM), and NIH grants K23-HL153888 (JEE) and DK062413 (DPBM).RESEARCH IN CONTEXTEvidence before this studyCurrently, several studies have shown the possible involvement of autoimmunity in patients affected by coronavirus disease 2019 (COVID-19). In contrast to cytokine storms, which tend to cause systemic, short-duration problems, autoantibodies (AABs) are thought to result in targeted, longer-term damage and development of autoimmune diseases.Added value of this studyAccording to our knowledge, we evaluated the largest number of protein antigens to characterize the prevalence and heterogeneity of the AABs signature in SARS-CoV-2 convalescent individuals. We examined autoimmune reactivity to SARS-CoV-2 in the absence of extreme clinical disease to acknowledge the existence of AABs even among those who had mild-to-moderate or no symptoms during their illness, as a hallmark of ongoing long-COVID syndrome. Through our analysis we suggest that VEGFA, MIF, IFNA4, SPP1 and APOH could be used as hallmark for SARS-CoV-2 infection and activation of the autoimmune system.Implications of all the available evidenceOur study comprehensively characterized the heterogeneity of the AABs signature in SARS-CoV-2 convalescent individuals. The results established a list of diagnostic signatures and potential therapeutic targets for long-Covid-19 patients although follow-up long-term studies are required. We believe that our findings will serve as a valuable resource, to drive further exploration of long-COVID syndrome pathogenesis.

BackgroundGiven the heterogeneity of clinical presentations, the diagnosis of Systemic Lupus Erythematosus (SLE) can be challenging, in particular in those patients presenting with early or incomplete disease, or with overlapping or atypical features. Autoantibodies (AABs) are important in aiding the clinical diagnosis of SLE, with some few AABs, anti-double-stranded DNA (dsDNA), anti-Smith (Sm), and anti-ribosomal P (riboP) being highly associated with SLE. As none of the traditional AABs has sufficient sensitivity to achieve diagnosis of SLE, current testing is based on measuring multiple AAB assays either in parallel or serial. We have recently identified novel AABs in SLE, which hold promise for improving diagnostic testing of SLE (1). We have developed quantitative ELISA-prototypes for five new AABs, which were tested in combination with traditional AABs.ObjectivesThe objectives of this study were to evaluate the diagnostic value of novel AABs and to screen for an optimized combination of novel and traditional AABs using logistic regression to increase the diagnostic accuracy of SLE testing.MethodsSerum samples were obtained from 156 SLE patients with European ancestry at the rheumatology department of the Heinrich-Heine University (Düsseldorf, Germany), and Hannover Medical School (Hannover, Germany). SLE samples were compared against 126 samples from autoimmune diseases (AID; myositis: n=20; Sjögren's syndrome (SjS): n=31; rheumatoid arthritis (RA) n=36; systemic sclerosis (SSc): n=39), and 77 healthy control samples. Prototype bead based ELISAs were developed for five novel antigens described in (1). Traditional diagnostic AABs were measured using IVD ELISAs and included: SSA/Ro60, SSA/Ro52, La/SSB, Sm, RNP, dsDNA, Scl70, CENPB, Jo-1, CCP, phospholipid and dsDNA. Optimized marker combinations of new and traditional markers were tested using logistic regression and receiver operating curve analysis (ROC).ResultsWhen comparing 156 SLE patients with 203 control samples, the area under the curve (AUC) of the five novel SLE ELISAs ranged from 0.63 to 0.75. A cut-off was set at a specificity of 95% and yielded a sensitivity ranging from 13.5% to 21.2% for the five novel assays. The sensitivity and specificity of new ELISAs was comparable to traditional ELISAs, which was in this cohort for anti-dsDNA 35% and 97%, anti-Sm 15% and 97%, and anti-RiboP 26% and 97%. A logistic regression model was used to combine the results of multiple tests. Compared to a logistic regression with traditional assays, a logistic regression with novel markers achieved higher sensitivity by pertaining high specificity. The logistic regression model based on a multimarker IVD assay with ten extracted nuclear antigens (ENA) yielded an AUC of 0.87 and a sensitivity of 58% at a specificity of 95%. By contrast, the optimal combination of traditional and novel ELISAs reached an AUC of 0.92 and a sensitivity of 75% at a specificity of 95%.ConclusionsThis study demonstrates the feasibility of combining test results of novel and traditional AABs using logistic regression to increase the diagnostic accuracy for SLE. Further studies are required to assess the impact of different ethnicities on marker selection and algorithm performance.ReferencesBudde P et al. (2016). Lupus. (8):812–22.Disclosure of InterestP. Budde Employee of: Protagen AG, H.-D. Zucht Employee of: Protagen AG, J. Schulte-Pelkum Employee of: Protagen AG, D. Wirtz Employee of: Protagen AG, P. Rengers Employee of: Protagen AG, T. Witte: None declared, P. Schulz-Knappe Employee of: Protagen AG, M. Schneider Consultant for: Protagen AG

BackgroundChronic inflammation is frequently observed on histological analysis of malignant and non-malignant prostate specimens. It is a suspected supporting factor for prostate diseases and their progression and a main cause of false positive PSA tests in cancer screening. We hypothesized that inflammation induces autoantibodies, which may be useful biomarkers. We aimed to identify and validate prostate inflammation associated serum autoantibodies in prostate cancer patients and evaluate the expression of corresponding autoantigens.MethodsRadical prostatectomy specimens of prostate cancer patients (N = 70) were classified into high and low inflammation groups according to the amount of tissue infiltrating lymphocytes. The corresponding pre-surgery blood serum samples were scrutinized for autoantibodies using a low-density protein array. Selected autoantigens were identified in prostate tissue and their expression pattern analyzed by immunohistochemistry and qPCR. The identified autoantibody profile was cross-checked in an independent sample set (N = 63) using the Luminex-bead protein array technology.ResultsProtein array screening identified 165 autoantibodies differentially abundant in the serum of high compared to low inflammation patients. The expression pattern of three corresponding antigens were established in benign and cancer tissue by immunohistochemistry and qPCR: SPAST (Spastin), STX18 (Syntaxin 18) and SPOP (speckle-type POZ protein). Of these, SPAST was significantly increased in prostate tissue with high inflammation. All three autoantigens were differentially expressed in primary and/or castration resistant prostate tumors when analyzed in an inflammation-independent tissue microarray. Cross-validation of the inflammation autoantibody profile on an independent sample set using a Luminex-bead protein array, retrieved 51 of the significantly discriminating autoantibodies. Three autoantibodies were significantly upregulated in both screens, MUT, RAB11B and CSRP2 (p>0.05), two, SPOP and ZNF671, close to statistical significance (p = 0.051 and 0.076).ConclusionsWe provide evidence of an inflammation-specific autoantibody profile and confirm the expression of corresponding autoantigens in prostate tissue. This supports evaluation of autoantibodies as non-invasive markers for prostate inflammation.