OpiCa1 Modulates Cardiomyocyte Viability Through PI3K/Akt Inhibition with Minimal Systemic Impact Beyond RyR Targeting

Introduction: Immune checkpoint inhibitors (ICIs) are associated with myocarditis; however, there is no consensus on strategies to identify or monitor for myocarditis. We sought to characterize longitudinal biomarkers in patients receiving ICIs. Methods: We conducted an observational cohort study of adult (≥18 years) patients who had received at least one dose of single or dual ICI at Michigan Medicine between June 2014 - December 2021. Patients were identified as having myocarditis if a clinical diagnosis was noted in the medical record by the cardiovascular consult team. Laboratory values (i.e., high-sensitivity troponin-T [hsTnT], aspartate transaminase [AST], alanine transaminase [ALT], lactate dehydrogenase [LDH], and creatinine phosphokinase [CPK]) were collected from medical records for 12 months after beginning ICI. Results: In 2,606 patients (mean (SD) age 64 (13) years; 60.8% male), 27 (1.0%) developed myocarditis at a median (IQR) time of 28 (18,40) days after starting ICIs. At diagnosis, most patients with myocarditis had a non-cardiac immune-related adverse event (88.9%) and an elevated ALT (88.9%), AST (85.2%), CPK (88.9%), and LDH (92.6%). All patients with myocarditis had an elevated hsTnT. Compared to patients without myocarditis, patients with myocarditis had significant elevations in ALT, AST, CPK, and LDH after starting ICIs (all P<0.001) (Figure). In patients with myocarditis, average concentrations of AST and ALT remained elevated at 12 months, whereas CPK peaked at 1 month then rapidly declined. Average concentrations of all biomarkers in patients without myocarditis were in the normal range. CPK values greater than the upper limit of normal had a sensitivity and specificity of detecting myocarditis of 99% and 23%, respectively. Conclusions: Myocarditis rarely occurs in the absence of other immune-related adverse events. Elevated AST, ALT and CPK should prompt measuring hs-TnT and initiating further evaluation for ICI myocarditis.

Transiently Elevated AST/LDH Are Associated with Clinical Response to Recombinant Circularly Permuted TRAIL (CPT) Plus Thalidomide in Patients with Relapsed and/or Refractory Multiple Myeloma

BackgroundIn March 2022, a concerning rise in cases of unexplained pediatric hepatitis was reported in multiple countries. Cases were defined as acute hepatitis with serum transaminases >500 U/L (aspartate transaminase [AST] or alanine transaminase [ALT]) in children aged 16 years or younger. We explored a simple federated data analytics method to search for evidence of unreported cases using routinely held data. We conducted a pragmatic survey to analyze changes in the proportion of hospitalized children with elevated AST or ALT over time. In addition, we studied the feasibility of using routinely collected clinical laboratory results to detect or follow-up the outbreak of an infectious disease.ObjectiveWe explored a simple federated data analytics method to search for evidence of unreported cases using routinely held data.MethodsWe provided hospitals with a simple computational tool to enable laboratories to share nondisclosive summary-level data. Summary statistics for AST and ALT measurements were collected from the last 10 years across all age groups. Measurements were considered elevated if ALT or AST was >200 U/L. The rate of elevated AST or ALT test for 3-week- to 5-year-olds was compared between a period of interest in which cases of hepatitis were reported (December 1, 2021, to August 31, 2022) and a prepandemic baseline period (January 1, 2012, to December 31, 2019). We calculated a z score, which measures the extent to which the rate for elevated ALT or AST was higher or lower in the period of interest compared to a baseline period, for the 3-week- to 5-year-olds.ResultsOur approach of sharing a simple software tool for local use enabled rapid, federated data analysis. A total of 34 hospitals in the United Kingdom, the Netherlands, Ireland, and Curaçao were asked to contribute summary data, and 30 (88%) submitted their data. For all locations combined, the rate of elevated AST or ALT measurements in the period of interest was not elevated (z score=−0.46; P=.64). Results from individual regions were discordant, with a higher rate of elevated AST or ALT values in the Netherlands (z score=4.48; P<.001), driven by results from a single center in Utrecht. We did not observe any clear indication of changes in primary care activity or test results in the same period.ConclusionsHospital laboratories collect large amounts of data on a daily basis that can potentially be of use for disease surveillance, but these are currently not optimally used. Federated analytics using nondisclosive, summary-level laboratory data sharing was successful, safe, and efficient. The approach holds potential as a tool for pandemic surveillance in future outbreaks. Our findings do not indicate the presence of a broader outbreak of mild hepatitis cases among young children, although there was an increase in elevated AST or ALT values locally in the Netherlands.

BackgroundOverall risks of hepatotoxicity with immune checkpoint inhibitors (ICIs) have yet to be compared in primary liver cancers to other solid tumors.MethodsWe reviewed data from the PubMed, Embase, and Scopus databases, and assessed the risk of hepatotoxicity associated with ICIs.ResultsA total of 117 trials were eligible for the meta‐analysis, including 7 trials with primary liver cancers. The most common hepatotoxicity was ALT elevation (incidence of all grade 5.29%, 95% CI 4.52-6.20) and AST elevation (incidence of all grade 5.88%, 95% CI 4.96-6.97). The incidence of all grade ALT and AST elevation was 6.01% and 6.84% for anti-PD‐1 (95% CI 5.04-7.18/5.69-8.25) and 3.60% and 3.72% for anti-PD-L1 (95% CI 2.72-4.76/2.82-4.94; p< 0.001/p<0.001). The incidence of ≥ grade 3 ALT and AST elevation was 1.54% and 1.48% for anti-PD‐1 (95% CI 1.19-1.58/1.07-2.04) and 1.03% and 1.08% for anti-PD-L1 (95% CI 0.71-1.51/0.80-1.45; p= 0.002/p<0.001). The incidence of all grade ALT and AST elevation was 13.3% and 14.2% in primary liver cancers (95% CI 11.1-16.0 and 9.93-20.36) vs. 4.92% and 5.38% in other solid tumors (95% CI 4.21-5.76 and 4.52-5.76 in other solid tumors; p <0.001/p<0.001).ConclusionOur study indicates that anti-PD-1 is associated with a higher risk of all‐ and high‐grade hepatotoxicity compared to anti-PD-L1, and primary liver cancers are associated with a higher risk of all‐ and high‐grade hepatotoxicity compared to other solid tumors.

Background: Hepatitis C is a common and important cause of chronic liver disease that often remains asymptomatic and most of the times discovered incidentally by routine serologic or biochemical testing. Aminotransferases (AST and ALT) reflect alterations in liver function/inflammation in liver diseases. The current study was conducted to examine the utilization and limitations of these biochemical markers in subjects with asymptomatic HCV infection. Aims and Objectives: To find out how many subjects with asymptomatic HCV infection have normal or elevated serum AST and ALT levels. Subjects and Methods: Study Design: Cross sectional. Study Duration: Seven months from November 2008 to July 2009. Study Universe: Services Hospital, Lahore. Study Population: This study included 413 subjects attending the outpatient departments of hospital with minor complaints. The random population of subjects was referred to the clinical laboratory of Services Hospital, Lahore for LFTs, HBsAg and anti-HCV screening from OPD department of the hospital. A total of 413 persons of all ages were analyzed during this study. There were 263 subjects who were sero – positive for anti-HCV (141 females, 122 males; median age 35 ± 11.34 years) and 150 in the control group (80 of them were females and 70 males with median age 32 ± 12.97 years) were sero-negative for both HBsAg and anti – HCV. Subjects Selection Criteria: In this study, only anti – HCV sero – positive subjects were included who was sero – negative for HBsAg or dual infection (both HBsAg and anti – HCV) and not on anti – viral therapy. Control group was sero – negative for both – HBsAg and anti – HCV antibodies. Data Collection: We assayed levels of serum aminotransferases (AST and ALT) and screened blood for HBsAg and anti-HCV. ELISA technique was used for viral hepatitis markers. Results: Out of 263 subjects tested positive for anti-HCV antibodies in their blood, 90.76 % and 87.45 % had elevated AST and ALT levels (ALT ≥ 40 U/L) respectively. Similarly, 9.23% and 12.54% had serum AST and ALT levels < 40 U/L. Fre-quency of elevated AST and ALT levels in individuals who were sero – negative for both infections (HBV or HCV) were 10.66% and 13.33%, respectively. 56.95% of anti – HCV antibody positive subjects had serum ALT elevation of less than two times the upper level of normal (ALT ≤ 80 U/L). Conclusion: The current study revealed that 90.76% and 87.45% had elevated AST and ALT levels (ALT ≥ 40 U/L) respec-tively. We concluded from this study that biochemical markers (AST, ALT and AST: ALT ratios) are useful, dependable and highly specific parameters for monitoring HCV infected patients (particularly asymptomatic) and frequent retesting is recom-mended to assess progression or regression of chronic liver disease. Key words: HCV infection, AST and ALT, clinical significance, prognosis.

INTRODUCTION: A significant number of subjects infected with COVID-19 present with gastrointestinal-related manifestations, particularly acute liver injury. Most data originate from China, and it is unclear that similar patterns hold in different ethno-social contexts. METHODS: A retrospective assessment of individuals diagnosed with COVID-19 in a county hospital in Minnesota was performed. Individuals with a positive COVID-19 test between 3/1/20 and 5/25/20 were included. We evaluated the role of race, ethnicity, co-morbidities, and proton-pump inhibitor (PPI) use on liver injury, hospital admission and mortality. Logistic regressions were performed using SAS 9.4. Statistical significance was reported at a 0.05 level. Census variables were sourced from the 2015 American Community Survey 5 year estimates. Results: 2164 individuals diagnosed with COVID-19 were identified. Median age was 39 years (IQR 28–51) and 52.4% were males. 12.5% were classified as white, 38.6% as African American, 34.6% as Hispanic, and 14.2% as other/unknown. Of those admitted to the hospital (N = 323), the median lab values were a follows: ALT 26 IU/L (IQR 18–45), AST 40 IU/L (IQR 27–63), total bilirubin 0.5 mg/dl (IQR 0.3–0.6) and alkaline phosphatase 74 IU/L (IQR 59–100). Only 2.6% and 3.7% of individuals without liver disease presented with elevated ALT or AST, respectively. In those with chronic liver disease (N = 71), 9.9% and 14.1% presented with elevated ALT or AST, respectively. Men were more likely to have abnormal ALT and AST (P = 0.002 and P = 0.005 respectively). Race was associated with an elevated AST on admission (P = 0.03) in those without chronic liver disease. We found no association between degree of abnormal liver enzymes and mortality. Surprisingly, being Hispanic or African American was inversely associated with mortality (P = 0.03 and P = 0.001, respectively). PPI use was associated with a 2.75 higher risk of mortality upon admission (P = 0.003) during bivariate analysis. However, this association became weak after correcting for age as well as renal, pulmonary and cardiovascular co-morbidities, suggesting that PPI-associated mortality could be related to other factors the lead to PPI use. CONCLUSION: Our findings suggest minimal hepatic injury inflicted by COVID-19 in those with no concurrent liver disease, regardless of race or ethnicity. Use of PPI was significantly associated to mortality, but further studies are needed to confirm this effect as the presence of co-morbidities significantly impacted this association.

5052 Background: EZN-4176 is a third generationLNA-ASO that binds the ligand binding domain of AR mRNA resulting in full length AR mRNA degradation and decreased AR protein expression. Methods: Patients (pts) (performance status ECOG≤1) with progressing CRPC were eligible; prior abiraterone and enzalutamide treatment were allowed. EZN-4176 was administered as a weekly (QW) one-hour intravenous infusion. The starting dose was 0.5 mg/Kg with a 4-week dose-limiting toxicity (DLT) period. After determination of the DLT and the maximum tolerated dose (MTD) for weekly administration, a fortnightly schedule (Q2W) was initiated; a 3+3 modified Fibonacci dose escalation design was pursued. PD studies evaluated AR expression in tissue utilizing antibodies to the amino and carboxy-termini of the AR. Results: 22 pts were enrolled (median age 70.6 years, range 59 – 84 years). One pt was treated with the Q2W schedule. Two DLTs (G3/G4 ALT/AST elevation) occurred at 10 mg/Kg, which was therefore identified as the MTD for the weekly schedule. Multiple pts treated at 6.5 and 10 mg/Kg (5/9 pts, 55%) developed ≥G2 ALT and/or AST elevation after the first cycle requiring dose reduction and treatment delay. The most frequent adverse events (AEs) all-grades were fatigue (21/22 pts, 95.4%), nausea (10/22 pts, 45.4%), constipation (8/22 pts, 36.3%), AST (8/22 pts, 36.3%) and ALT (10/22 pts, 45.4%) elevation. The most frequent G3/4 AEs were AST (4/22 pts, 18.1%) and ALT (5/22 pts, 22.7%) elevation. Maximum PSA and circulating tumor cells (CTCs) declines are summarized below. There were no objective soft tissue responses. PD studies did not document any knockdown of AR expression Conclusions: EZN-4176 has limited antitumour activity in CRPC at its MTD for weekly administration. Safety, PK, PD and efficacy data will be presented. Clinical trial information: NCT01337518. [Table: see text]

209 Background: Hormonal therapies function through CDK4/CDK6/E2F axis which is essential in luminal estrogen receptor positive breast cancer. Reactivation of these CDK4 and CDK6 kinases has been implicated in endocrine resistance. Many CDK 4/6 inhibitors were employed in studies with noteworthy safety concerns. We performed a systematic review and meta-analysis of randomized controlled trials (RCTs) to determine the risk of gastrointestinal (GI) and hepatic toxicities. Methods: MEDLINE, EMBASE databases and meeting abstracts from inception through June 2017 were queried. RCTs that mention GI symptoms and elevation of liver enzymes as adverse effects were incorporated in the analysis. Mantel-Haenszel method was used to calculate the estimated pooled risk ratio with 95% confidence interval (CI). Results: We included five RCTs with a total of 2671 patients treated with CDK 4/6 inhibitors. The study arm used palbociclib-letrozole, palbociclib-fulvestrant, ribociclib-letrozole and abemaciclib-fulvestrant while the control arm used placebo in combination with letrozole or fulvestrant. The relative risks (RR) of all-grade side effects were as follows: diarrhea, 1.72 (95% CI: 1.08 – 2.74, p = 0.02); stomatitis, 2.62 (95% CI: 1.43 – 4.79, p = 0.002); nausea, 1.59 (95% CI: 1.30–1.94; p &lt; 0.0001); vomiting, 1.65 (95% CI: 1.06–2.56; p = 0.02); elevated AST, 2.30 (95% CI: 1.34–3.93; p = 0.002); and elevated ALT, 2.78 (95% CI: 1.90–4.08; p &lt; 0.0001). The RR of high-grade side effects were as follows: diarrhea, 2.26 (95% CI: 0.39 – 13.09, p = 0.36); stomatitis, 2.14 (95% CI: 0.36 – 12.72, p = 0.40); nausea, 1.13 (95% CI: 0.28 –4.47; p = 0.86); vomiting, 0.79 (95% CI: 0.24 – 2.54; p = 0.69); elevated AST, 1.85 (95% CI: 0.74 – 4.64; p = 0.18); and elevated ALT, 4.33 (95% CI: 2.15 – 8.71; p &lt; 0.0001). Conclusions: The risk of developing any-grade diarrhea, stomatitis, nausea, vomiting and elevated AST and ALT, was high in CDK 4/6 inhibitors based regimen. Moreover, it increased the risk of high-grade elevated ALT.

The objective was to assess the effects of commercial regular diet as control, total food restriction with honey, commercial regular diet with dextrose, or total food restriction with dextrose, on blood variables after carbon tetrachloride (CCl4) administration. Sprague Dawley albino rats were divided into four groups, 10 rats each; Group 1 rats were on commercial regular diet, Group 2 rats were on commercial regular diet with 50% dextrose, Group 3 rats were on total food restriction with 50% dextrose, and Group 4 rats were on total food restriction with 50% honey. Rats in all the groups were i.m. administered CCL4 (2.4 mL kg b. wt.−1). Blood tests including ALT, AST, serum albumin, serum protein, BUN, blood glucose (BG), hemoglobin (Hb), and white blood cell (WBC) were performed before CCl4 administration and repeated after 48 and 96 h of post-injection. In Group 1, CCl4 caused significant elevation in AST and ALT, and decrease in BS, WBC, and BUN; lower elevation in AST and ALT at 48 h and decreased AST and ALT at 96 h were obtained when dextrose was added to commercial regular diet (Group 2). Using dextrose alone (Group 3), though there was significant elevation of AST and ALT and decrease in BUN and WBC as compared to baseline values, significant decrease in ALT, AST, and BUN as compared to control was obtained. During absolute honey feeding (Group 4), elevation in AST and ALT obtained, following CCl4 administration was significantly less than the values obtained in all other groups; with lower elevation in AST and ALT as compared to baseline values. Honey increased serum albumin, serum protein, BG, and caused lower reduction in Hb. Conclusively, exclusive honey feeding (50% concentration) significantly modifies and ameliorates biochemical and hematological changes obtained after CCl4 injection.

BackgroundThe use of pyronaridine-artesunate (PA) has been associated with scarce transaminitis in patients. This analysis aimed to evaluate the hepatic safety profile of repeated treatment with PA versus artemether–lumefantrine (AL) in patients with consecutive uncomplicated malaria episodes in Bobo-Dioulasso, Burkina Faso.MethodsThis study analysed data from a clinical trial conducted from 2012 to 2015, in which participants with uncomplicated malaria were assigned to either PA or AL arms and followed up to 42 days. Subsequent malaria episodes within a 2-years follow up period were also treated with the same ACT initially allocated. Transaminases (AST/ALT), alkaline phosphatase (ALP), total and direct bilirubin were measured at days 0 (baseline), 3, 7, 28 and on some unscheduled days if required. The proportions of non-clinical hepatic adverse events (AEs) following first and repeated treatments with PA and AL were compared within study arms. The association of these AEs with retreatment in each arm was also determined using a logistic regression model.ResultsA total of 1379 malaria episodes were included in the intention to treat analysis with 60% of all cases occurring in the AL arm. Overall, 179 non-clinical hepatic AEs were recorded in the AL arm versus 145 in the PA arm. Elevated ALT was noted in 3.05% of treated malaria episodes, elevated AST 3.34%, elevated ALP 1.81%, and elevated total and direct bilirubin in 7.90% and 7.40% respectively. Retreated participants were less likely to experience elevated ALT and AST than first episode treated participants in both arms. One case of Hy’s law condition was recorded in a first treated participant of the PA arm. Participants from the retreatment group were 76% and 84% less likely to have elevated ALT and AST, respectively, in the AL arm and 68% less likely to present elevated ALT in the PA arm. In contrast, they were almost 2 times more likely to experience elevated total bilirubin in both arms.ConclusionsPyronaridine-artesunate and artemether–lumefantrine showed similar hepatic safety when used repeatedly in participants with uncomplicated malaria. Pyronaridine-artesunate represents therefore a suitable alternative to the current first line anti-malarial drugs in use in endemic areas.Trial registration Pan African Clinical Trials Registry. PACTR201105000286876

Myocardial ischemia and reperfusion occurs in myocardial infarction. Timely reperfusion will exacerbate the injury through the mitochondria-mediated apoptosis in cardiomyocytes due to the accumulation of excessive reactive oxygen species (ROS). In order to identify novel therapeutic approaches, the cardioprotective effects of carnosic acid and the underlying mechanisms were investigated in the present study in H9c2 cardiomyocytes injured by hypoxia/reoxygenation in vitro. The viability of H9c2 cardiomyocytes was detected by MTT assay and further confirmed by the detection of intracellular lactate dehydrogenase (LDH) release. The mitochondrial function in H9c2 cardiomyocytes was evaluated using fluorescence methods. The proteins related to apoptosis, including caspase-3, B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were analyzed by western blot analysis, and the activity of caspase-3 was determined using a colorimetric method. As a result, carnosic acid was demonstrated to improve the viability of H9c2 cardiomyocytes and suppress the leakage of cytosolic lactate dehydrogenase under hypoxia/reoxygenation. In addition, the overproduction of intracellular ROS and intracellular calcium overload were ameliorated in the presence of carnosic acid. The dysfunction of mitochondria in H9c2 cardiomyocytes was also attenuated by carnosic acid through blocking the collapse of mitochondrial membrane potential (MMP) and mitochondrial permeability transition pore (mPTP) opening. Furthermore, the apoptosis of H9c2 cardiomyocytes resulted from hypoxia/reoxygenation was inhibited by carnosic acid through the upregulation of Bcl-2 and the downregulation of Bax and caspase-3 levels. These results provided evidence for further investigation that would assist in the development of novel therapeutic approaches for myocardial infarction.

Background. Differentiation among types of muscular dystrophy (MD) has remained challenging. In this retrospective study, we sought to develop a methodology for differentiation of MD types using analysis of serum enzyme profiles. Methods. The serum levels of enzymes from 232 patients, including 120 with DMD, 36 with BMD, 36 with FSHD, 46 with LGMD, and 11 with EDMD, were evaluated. Results. The characteristic profiles of serum enzymes facilitated differentiation of these five types of MD. DMD was characterized by simultaneous elevation of ALT, AST, LDH, and ALP; BMD and LGMD were characterized by elevation of ALT, AST, and LDH; and FSHD and EDMD were characterized by a lack of abnormal serum enzyme levels. We further developed discriminant functions to distinguish BMD and LGMD. For LGMD, LGMD2B patients had significantly higher ALP levels than non-LGMD2B patients (98 ± 59 U/L versus 45 ± 9 U/L, resp., p < 0.05). Conclusions. Our approach enabled the determination of MD subtypes using serum enzyme profiles prior to genetic testing, which will increase the chance a mutation will be found in the first gene analyzed.

Consolidation with Nivolumab and Brentuximab Vedotin after Autologous Hematopoietic Cell Transplantation in Patients with High-Risk Hodgkin Lymphoma

Results from Ongoing Phase 1/2 Trial of SL-401 As Consolidation Therapy in Patients with Acute Myeloid Leukemia (AML) in Remission with Minimal Residual Disease (MRD)

Abnormal aminotransferase activity in women with polycystic ovary syndrome