PBC Patients Research Articles

SARS-CoV-2 vaccination response in patients with autoimmune hepatitis and autoimmune cholestatic liver disease.

Background/AimsIn this observational study, we explored the humoral and cellular immune response to SARS‐CoV‐2 vaccination in patients with autoimmune hepatitis (AIH) and patients with cholestatic autoimmune liver disease (primary sclerosing cholangitis [PSC] and primary biliary cholangitis [PBC]).MethodsAnti‐SARS‐CoV‐2 antibody titers were determined using the DiaSorin LIAISON and Roche immunoassays in 103 AIH, 64 PSC, and 61 PBC patients and 95 healthy controls >14 days after the second COVID‐19 vaccination. The spike‐specific T‐cell response was assessed using an activation‐induced marker assay (AIM) in a subset of individuals.ResultsPrevious SARS‐CoV‐2 infection was frequently detected in AIH but not in PBC/PSC (10/112 (9%), versus 4/144 (2.7%), p = 0.03). In the remaining patients, seroconversion was measurable in 97% of AIH and 99% of PBC/PSC patients, respectively. However, in 13/94 AIH patients antibody levels were lower than in any healthy control, which contributed to lower antibody levels of the total AIH cohort when compared to PBC/PSC or controls (641 vs. 1020 vs. 1200 BAU/ml, respectively). Notably, antibody levels were comparably low in AIH patients with (n = 85) and without immunosuppression (n = 9). Also, antibody titers significantly declined within 7 months after the second vaccination. In the AIM assay of 20 AIH patients, a spike‐specific T‐cell response was undetectable in 45% despite a positive serology, while 87% (13/15) of the PBC/PSC demonstrated a spike‐specific T‐cell response.ConclusionPatients with AIH show an increased SARS‐CoV‐2 infection rate as well as an impaired B‐ and T‐cell response to SARS‐CoV‐2 vaccine compared to PBC and PSC patients, even in the absence of immunosuppression. Thus, antibody responses to vaccination in AIH patients need to be monitored and early booster immunizations considered in low responders.

Effect of LncRNA XIST on Immune Cells of Primary Biliary Cholangitis.

ObjectivePrimary biliary cholangitis (PBC) is an autoimmune disease with significant gender difference. X chromosome inactivation (XCI) plays important roles in susceptibility to diseases between genders. This work focuses on the differences of LncRNA XIST in several defined immune cells populations as well as its effects on naive CD4+ T cells proliferation and differentiation in patients with PBC.MethodsNKs, B cells, CD4+ T, and CD8+ T cells were separated by MicroBeads from peripheral blood mononuclear cells (PBMCs) of PBC patients and healthy control (HC). The expression levels of LncRNA XIST in these immune cells were quantified by qRT-PCR and their subcellular localized analyzed by FISH. Lentivirus were used to interfere the expression of LncRNA XIST, and CCK8 was used to detect the proliferation of naive CD4+ T cells in PBC patients. Finally, naive CD4+ T cells were co-cultured with the bile duct epithelial cells (BECs), and the effects of LncRNA XIST on the typing of naive CD4+ T cells and related cytokines were determined by qRT-PCR and ELISA.ResultsThe expression levels of LncRNA XIST in NKs and CD4+ T cells in PBC patients were significantly higher than those in HC, and were primarily located at the nucleus. LncRNA XIST could promote the proliferation of naive CD4+ T cells. When naive CD4+ T cells were co-cultured with BECs, the expressions of IFN-γ, IL-17, T-bet and RORγt in naive CD4+ T cells were decreased.ConclusionLncRNA XIST was associated with lymphocyte abnormalities in patients with PBC. The high expression of LncRNA XIST could stimulate proliferation and differentiation of naive CD4+ T cells, which might account for the high occurrence of PBC in female.

Abstract P3-16-04: Estimation of willingness-to-pay for breast cancer treatments through contingent valuation method in Japanese breast cancer patients (JCOG1709A): The main study findings

Abstract Background In 2019, the Japanese government has introduced the economic valuation to healthcare system. However, the threshold for determining cost-effectiveness-analysis has yet to be adequately studied. Therefore, we have designed a study to examine the financial value of “life” and “health” based on willingness-to-pay (WTP) considered by patients with primary and metastatic breast cancer (PBC and MBC) in Japan. Methods This study is conducted by forty-five hospitals under the Breast Cancer Study Group of Japan Clinical Oncology Group (JCOG). In the preliminary study, 168 patients (84 patients with PBC and 84 patients with MBC) from 20 to 79 years of age paying medical costs were examined their WTP for setting up the dichotomous-choice method survey form for using in the main study. Virtual scenario-specific treatments to avoid the recurrence and death of breast cancer for one year for PBC patients, and to prolong the survival period for one year for the patients with MBC were presented. The patients were evaluated how much money would pay to receive the treatment in a self-written answer and the amount of WTP that would be presented in the main study was determined. In the main study, 1,620 patients (810 patients with PBC and 810 patients with MBC) were surveyed to their WTP for hypothetical scenarios using the dichotomous-choice method survey form. With the two-step dichotomous-choice method, if the first stage response was yes, a higher amount was displayed for the second stage. If the first stage response was no, a lower amount was shown for the second stage. For the main analysis, the acceptance probability curve was calculated using the parametric method. The calculation was based on the obtained responses and median WTP, which was determined by calculating the amount at which the acceptance probability curve is at 50%. The obtained amount was converted to WTP per one QALY based on the displayed question content. Results Completed surveys were collected from 716 PBC patients (88.4%) and 702 patients (86.7%). Based on the preliminary study, the amount of WTP offering in the first stage was set 5 million yen (JPY) (50,000 USD), and offering in the second stage was 7 million JPY (70,000 USD) for those who responded accepting the scenario treatment and 3 million JPY (30,000 USD) for those who responded refusing. We obtained point estimates of WTP for each virtual scenario treatments. The WTP of the two hypothetical scenarios for PBC patients were as follows; scenario 1) the treatment to gain one year without recurrence: 2.06 million JPY (20,600 USD), scenario 2) the treatment that reduces risk of recurrence by 40%: 2.40 million JPY (24,000 USD). The WTP of the two hypothetical scenarios for MBC patients were as follows; scenario 3) the treatment that can prolong survival for one year in current health condition: 2.81 million JPY (28,100 USD), scenario 4) the treatment that can prolong survival by one year in the health condition before having breast cancer: 2.92 million JPY (29,200 USD). Conclusion We created hypothetical 1-quality-adjusted survival year (QALY) scenarios for breast cancer patients with different disease status and validated them using the contingent valuation method. Our results demonstrate that WTP per 1QALY varies with the disease status of breast cancer. Therefore, we conclude that we cannot simply use a uniform threshold when examining the cost-effectiveness of medical interventions in breast cancer as a unit of cost/QALY. Citation Format: Tsuguo Iwatani, Fumikata Hara, Tadahiko Shien, Masato Takahashi, Norikazu Masuda, Yasuaki Sagara, Keita Sasaki, Junki Mizusawa, Haruhiko Fukuda, Takaru Shiroiwa, Hiroji Iwata. Estimation of willingness-to-pay for breast cancer treatments through contingent valuation method in Japanese breast cancer patients (JCOG1709A): The main study findings [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-16-04.

Left ventricular systolic function parameters are increased in patients with primary biliary cholangitis without cirrhosis

Abstract Funding Acknowledgements Type of funding sources: None. Background Primary biliary cholangitis (PBC) is a chronic, autoimmune liver disease causing destruction of the small intralobular bile ducts in the liver. PBC patients can also develop extrahepatic manifestations, mostly related to skin, nervous system and laboratory abnormalities. Cardiac involvement is inconsistent in PBC studies despite high levels of serum lipids. Cardiac abnormalities are especially important because with the treatment many patients with early-stage PBC may have a normal life expectancy and therefore the cardiovascular complications can develop. Additionally, in recent years new echocardiographic modalities emerged letting for more precise analysis of cardiac morphology and function. Purpose The purpose of the study was to compare novel and classic echocardiographic indices in patients with confirmed PBC without cirrhosis and in control subjects. Methods 22 female patients with PBC and without cirrhosis and 44 healthy females, matched for age, (53.3 ± 9 years and 50.9 ± 8 years respectively, p = 0.065) were investigated. TTE study was performed and analysis of left ventricular (LV) global longitudinal strain (GLS) and myocardial work was performed. Results Systolic blood pressure (SBP) differed between PBC and control group (128 ± 17 mmHg and 116 ± 11 mmHg respectively, p = 0.006), while diastolic blood pressure (DBP) (78 ± 10 mmHg and 76 ± 10 mmHg, p = 0.2) and heart rate (HR) (73 ± 12 bpm and 70 ± 8 bpm , p = 0.32) did not. PBC group had increased diameter of cardiac chambers when adjusted for body surface area (BSA), including left atrium (LA/BSA) and right ventricle (RV/BSA), as well as left ventricular mass (LWMI) and relative wall thickness (RWT) (Table) with no difference in left ventricular end diastolic diameter (LVEDD/BSA). LV ejection fraction (EF) measured by Simpson method was increased in PBC group (Table). LV diastolic function parameters (E/A, E/e’) were impaired in patients with PBC when compared with controls (Table 1). LV GLS indices (GLSAVG, GLSLAX, GLS4C, GLS2C) were significantly higher in PBC group, as well as global myocardial work indexes : global myocardial work (GWI) and global constructive work (GCW). Global work efficiency (GWE) and global wasted work (GWW) indexes were not different (Table). Conclusion Our study shows that in patients with PBC without cirrhosis LV systolic function is increased. That implies hyperkinetic circulation in that group of patients even before the development of cirrhosis. Abstract Table

PRIMARY BILIARY CHOLANGITIS COMPLICATED WITH ULCERATIVE COLITIS: A CASE REPORT

Liver involvement is not unusual in patients with inflammatory bowel disease (IBD), where one third of patients have abnormal liver biochemical tests, becoming a diagnostic challenge. Primary biliary cholangitis (CPB) is an autoimmune liver disease that presents with chronic cholestasis, the presence of specific antibodies and histological findings of destructive non-suppurative cholangitis. Genetic, immunological, and environmental factors that contribute to the pathogenesis of IBD may also contribute to associated hepatobiliary disorders. Objective: Present the case of a 67-year-old woman who consulted for cholestatic symptoms. Medical record of smoking for 25 years was suspended 15 years ago with a smoking index of 5. She reported a history of weakness, asthenia, self-limited palmar and plantar pruritus of 2 years of evolution, associated with abnormal liver function tests (total bilirubin 1.8 mg / dl, INR 1.40, albumin 3.2 g / dL). The initial physical examination revealed jaundice in the sclera, palmar erythema, evidence of telangiectasia in the abdomen, collateral circulation with medusa caput, positive ascitic wave, and splenomegaly. R factor is cholestatic, with alkaline phosphatase of 416 U / L, gamma glutamyl transpeptidase 660 U / L, alanine transferase 59 U / L, for which possible viral and autoimmune causes are addressed as the first possibility, reflecting antimitochondrial antibodies with high titers (278 U / L), associated with immunoglobulin G of 2490 mg / dl and immunoglobulin M of 734 mg / dl. During her one-year follow-up, she reported the onset of diarrheal stools Bristol 6, 2 to 3 episodes per day, with occasional urgency and with inflammatory characteristics, due to the presence of mucus and blood. Infectious causes are ruled out, elevated acute phase reactants are reported, followed up with a colonoscopy and biopsy samples, reporting the presence of nonspecific proctitis, chronic colitis with focal ulceration, lymphoid aggregates, focal atrophy, and glandular distortion compatible with ulcerative colitis (UC). (Panel A) Due to the unusual association between UC and PBC, magnetic resonance cholangiography was requested, ruling out the overlap syndrome between primary biliary cholangitis and primary sclerosing cholangitis. (Panel B) A diverse heterogeneous group of hepatobiliary manifestations is reported in both UC and CD, and approximately 5% of adults with IBD have developed chronic liver disease. PBC is not usually associated with IBD, and concomitant reported cases are anecdotal. The presentations are different than the typical CBP without UC. PBC usually affects middle-aged women. The sex ratio is 10: 1 (female to male) and the mean age at diagnosis is 57.5 years. While the disease tends to affect men more often, with a female / male sex ratio of 2: 1 when associated with IBD. The distribution of ulcerative colitis in PBC patients is usually mild with limited bowel involvement. In a review by Tasa et al., eleven of 15 patients described left side colitis and proctitis. The association between PBC and UC remains rare, as there are still few reported cases regarding the combined presentation of these diseases. Although PSC is the most specific hepatobiliary manifestation among UC patients with cholestasis, PBC should be considered in those with unexplained intrahepatic cholestasis. The use of a reliable test such as AMAs is of utmost importance to avoid misdiagnosis and/or under diagnosis. The authors declare that there is no conflict of interest.

Distinct prognostic value of different portal hypertension-associated features in patients with primary biliary cholangitis

BackgroundPrimary biliary cholangitis (PBC) may progress to cirrhosis and clinically significant portal hypertension (CSPH). This study assesses different features of CSPH and their distinct prognostic impact regarding decompensation and survival in patients with PBC.MethodsPatients with PBC were identified during a database query of our digital patient reporting system.ResultsA total of 333 PBC patients (mean age 54.3 years, 86.8% females, median follow-up 5.8 years) were retrospectively assessed and 127 (38.1%) showed features of CSPH: 63 (18.9%) developed varices, 98 (29.4%) splenomegaly, 62 (18.6%) ascites and 20 (15.7%) experienced acute variceal bleeding. Splenomegaly, portosystemic collaterals and esophageal varices were associated with an increased 5-year (5Y) risk of decompensation (15.0%, 17.8% and 20.9%, respectively). Patients without advanced chronic liver disease (ACLD) had a similar 5Y-transplant free survival (TFS) (96.6%) compared to patients with compensated ACLD (cACLD) but without CSPH (96.9%). On the contrary, PBC patients with cACLD and CSPH (57.4%) or decompensated ACLD (dACLD) (36.4%) had significantly decreased 5Y survival rates. The combination of LSM < 15 kPa and platelets ≥ 150G/L indicated a negligible risk for decompensation (5Y 0.0%) and for mortality (5Y 0.0%). Overall, 44 (13.2%) patients died, with 18 (40.9%) deaths attributed to CSPH-related complications.ConclusionIn PBC, features of CSPH may occur early and indicate an increased risk for subsequent decompensation and mortality. Hence, regular screening and on-time treatment for CSPH is crucial. Combining LSM and platelets serves as a valuable preliminary assessment, as LSM < 15 kPa and platelets ≥ 150G/L indicate an excellent long-term outcome.

Evaluation of headaches in primary brain calcification in Japan

AbstractBackgroundPrimary brain calcification (PBC) is a rare neurodegenerative disorder characterized by bilateral calcification in the brain. No nationwide survey on headache has been performed in Japan.AimA questionnaire survey was performed to clarify the situation of headache in PBC patients.MethodsThe first questionnaire survey was conducted on 173 patients diagnosed as having PBC in Japan. Subsequently, the second survey was performed using two types of questionnaire: Headache Impact Test (HIT‐6) and Migraine Disability Assessment (MIDAS).ResultsIn the first survey, 27 of 82 patients had headaches. Only one patient was able to be classified in accordance with the criteria as having migraine without aura. In the second survey, 10 of 17 patients showed a significant score of 50 and higher in HIT‐6, and five of 17 patients showed a significant score of 6 and higher in MIDAS. No significant correlation was found between total calcification score and HIT‐6 score or MIDAS score in the second survey. All patients carrying the PDGFB variant and two of three patients carrying the SLC20A2 variant showed some effects of headache on their daily life as judged from their MIDAS scores or HIT‐6 score.ConclusionsThe headaches in PBC patients could not be classified as the primary headache. Those headaches in PBC patients may be of the secondary headache caused by the dysfunction of the blood‐brain barrier. This first report on headaches of Japanese patients with PBC will contribute to the understanding of PBC patients' difficulties in their daily lives.

Interleukin 23 Produced by Hepatic Monocyte-Derived Macrophages Is Essential for the Development of Murine Primary Biliary Cholangitis.

Background and AimsPrimary Biliary Cholangitis (PBC) is an organ-specific autoimmune liver disease. Mononuclear phagocytes (MNPs), comprise of monocyte, dendritic cells and monocyte-derived macrophages, constitute major arm of the innate immune system known to be involved in the pathogenesis of autoimmune disorders. MNPs were shown to accumulate around intra-hepatic bile ducts in livers of PBC patients. Interleukin 23 (IL-23) is a pro-inflammatory cytokine. IL-23-positive cells were detected in livers of patients with advanced stage PBC and IL-23 serum levels found to be in correlation with PBC disease severity. Our overall goal was to assess the importance of IL-23 derived from MNPs in PBC pathogenesis.MethodsWe utilized an inducible murine model of PBC and took advantage of transgenic mice targeting expression of IL-23 by specific MNP populations. Analysis included liver histology assessment, flow cytometry of hepatic immune cells and hepatic cytokine profile evaluation. Specific MNPs sub-populations were sorted and assessed for IL-23 expression levels.ResultsFlow cytometry analysis of non-parenchymal liver cells in autoimmune cholangitis revealed massive infiltration of the liver by MNPs and neutrophils and a decrease in Kupffer cells numbers. In addition, a 4-fold increase in the incidence of hepatic IL-17A producing CD4+ T cells was found to be associated with an increase in hepatic IL23-p19 and IL17A expression levels. Disease severity was significantly ameliorated in both CD11ccreP19flox/flox and CX3CR1creP19 flox/flox mice as assessed by reduced portal inflammation and decreased hepatic expression of various inflammatory cytokines. Amelioration of disease severity was associated with reduction in IL-17A producing CD4+ T cells percentages and decreased hepatic IL23-p19 and IL17A expression levels. qRT-PCR analysis of sorted hepatic MNPs demonstrated high expression levels of IL-23 mRNA specifically by CX3CR1hiCD11c+ monocyte-derived macrophages.ConclusionOur results indicate a major role for IL-23 produced by hepatic monocyte-derived macrophages in the pathogenesis of PBC. These results may pave the road for the development of new immune-based and cell specific therapeutic modalities for PBC patients not responding to current therapies.

Changes of Serum IgG Glycosylation Patterns in Primary Biliary Cholangitis Patients.

ObjectivePrimary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease whose diagnosis is based significantly on autoantibody detection. This study aims to investigate the glycosylation profile of serum IgG in PBC patients using high-throughput lectin microarrays technology.MethodLectin microarray containing 56 lectins was used to detect and analyze the expression of serum IgG glycosylation in 99 PBC patients, 70 disease controls (DCs), and 38 healthy controls (HCs). Significant differences in PBC from control groups as well as across PBC subgroups positive for various autoantibodies were explored and verified by lectin blot technique.ResultsLectin microarray detection revealed that compared to DC and HC groups, the specific glycan level of serum IgG sialic acid in PBC patients was increased. For each PBC subgroup, glycan levels of IgG mannose and galactose were decreased in AMA-M2 positive PBC patients compared to the AMA-M2 negative group. IgG N-Acetylgalactosamine (GalNAc) and fucose were decreased in anti-sp100 positive patients. IgG galactose was increased in anti-gp210 positive patients. IgG mannose was decreased in ACA-positive patients. Although the difference in overall sialic acid level was not observed using lectin blot, all results among the above PBC subgroups were consistent with the results of the technique.ConclusionLectin microarray is an effective and reliable technique for analyzing glycan structure. PBC patients positive for different autoantibody exhibits distinct glycan profile. Altered levels of glycosylation may be related to the occurrence and development of the disease, which could provide a direction for new biomarker identification.

Prognostic Factors in Primary Biliary Cholangitis: A Retrospective Study of Joint Slovak and Croatian Cohort of 249 Patients.

Objective: To identify pretreatment laboratory parameters associated with treatment response and to describe the relationship between treatment response and liver decompensation in patients with primary biliary cholangitis treated with ursodeoxycholic acid. Methods: We defined treatment response as both ALP ≤ 1.67 × ULN and total bilirubin ≤ 2 × ULN. Multiple logistic regression analyses were performed to adjust for confounding effects of sociodemographic variables. Results: Pretreatment total bilirubin ((TB); OR = 0.3388, 95%CI = 0.1671–0.6077), ALT (OR = 0.5306, 95%CI = 0.3830–0.7080), AST (OR = 0.4065, 95%CI = 0.2690–0.5834), ALP (OR = 0.3440, 95%CI = 0.2356–0.4723), total cholesterol ((TC); OR = 0.7730, 95%CI = 0.6242–0.9271), APRI (OR = 0.3375, 95%CI = 0.1833–0.5774), as well as pretreatment albumin (OR = 1.1612, 95%CI = 1.0706–1.2688) and ALT/ALP (OR = 2.4596, 95%CI = 1.2095–5.5472) were associated with treatment response after six months of treatment. Pretreatment TB (OR = 0.2777, 95%CI = 0.1288–0.5228), ALT (OR = 0.5968, 95%CI = 0.4354–0.7963), AST (OR = 0.4161, 95%CI = 0.2736–0.6076), ALP (OR = 0.4676, 95%CI = 0.3487–0.6048), APRI (OR = 0.2838, 95%CI = 0.1433–0.5141), as well as pretreatment albumin (OR = 1.2359, 95%CI = 1.1257–1.3714) and platelet count (OR = 1.0056, 95%CI = 1.0011–1.0103) were associated with treatment response after 12 months of treatment. Treatment response after 6 months of UDCA therapy is significantly associated with treatment response after 12 months of UDCA therapy (OR = 25.2976, 95% CI = 10.5881–68.4917). Treatment responses after 6 and 12 months of UDCA therapy decrease the risk of an episode of liver decompensation in PBC patients (OR = 12.1156, 95%CI = 3.7192–54.4826 and OR = 21.6000, 95%CI = 6.6319–97.3840, respectively). Conclusions: There are several pretreatment laboratory parameters associated with treatment response in patients with primary biliary cholangitis. Treatment response after six months is significantly associated with treatment response after 12 months of ursodeoxycholic acid (UDCA) therapy. Treatment responses after 6 and 12 months of UDCA decrease the risk of an episode of liver decompensation.

S100 calcium binding protein A6 and associated long noncoding ribonucleic acids as biomarkers in the diagnosis and staging of primary biliary cholangitis.

BACKGROUNDPrimary biliary cholangitis (PBC) is a chronic and slowly progressing cholestatic disease, which causes damage to the small intrahepatic bile duct by immuno-regulation, and may lead to cholestasis, liver fibrosis, cirrhosis and, eventually, liver failure.AIMTo explore the potential diagnosis and staging value of plasma S100 calcium binding protein A6 (S100A6) messenger ribonucleic acid (mRNA), LINC00312, LINC00472, and LINC01257 in primary biliary cholangitis.METHODSA total of 145 PBC patients and 110 healthy controls (HCs) were enrolled. Among them, 80 PBC patients and 60 HCs were used as the training set, and 65 PBC patients and 50 HCs were used as the validation set. The relative expression levels of plasma S100A6 mRNA, long noncoding ribonucleic acids LINC00312, LINC00472 and LINC01257 were analyzed using quantitative reverse transcription-polymerase chain reaction. The bile duct ligation (BDL) mouse model was used to simulate PBC. Then double immunofluorescence was conducted to verify the overexpression of S100A6 protein in intrahepatic bile duct cells of BDL mice. Human intrahepatic biliary epithelial cells were treated with glycochenodeoxycholate to simulate the cholestatic environment of intrahepatic biliary epithelial cells in PBC.RESULTSThe expression of S100A6 protein in intrahepatic bile duct cells was up-regulated in the BDL mouse model compared with sham mice. The relative expression levels of plasma S100A6 mRNA, log10 LINC00472 and LINC01257 were up-regulated while LINC00312 was down-regulated in plasma of PBC patients compared with HCs (3.01 ± 1.04 vs 2.09 ± 0.87, P < 0.0001; 2.46 ± 1.03 vs 1.77 ± 0.84, P < 0.0001; 3.49 ± 1.64 vs 2.37 ± 0.96, P < 0.0001; 1.70 ± 0.33 vs 2.07 ± 0.53, P < 0.0001, respectively). The relative expression levels of S100A6 mRNA, LINC00472 and LINC01257 were up-regulated and LINC00312 was down-regulated in human intrahepatic biliary epithelial cells treated with glycochenodeoxycholate compared with control (2.97 ± 0.43 vs 1.09 ± 0.08, P = 0.0018; 2.70 ± 0.26 vs 1.10 ± 0.10, P = 0.0006; 2.23 ± 0.21 vs 1.10 ± 0.10, P = 0.0011; 1.20 ± 0.04 vs 3.03 ± 0.15, P < 0.0001, respectively). The mean expression of S100A6 in the advanced stage (III and IV) of PBC was up-regulated compared to that in HCs and the early stage (II) (3.38 ± 0.71 vs 2.09 ± 0.87, P < 0.0001; 3.38 ± 0.71 vs 2.57 ± 1.21, P = 0.0003, respectively); and in the early stage (II), it was higher than that in HCs (2.57 ± 1.21 vs 2.09 ± 0.87, P = 0.03). The mean expression of LINC00312 in the advanced stage was lower than that in the early stage and HCs (1.39 ± 0.29 vs 1.56 ± 0.33, P = 0.01; 1.39 ± 0.29 vs 2.07 ± 0.53, P < 0.0001, respectively); in addition, the mean expression of LINC00312 in the early stage was lower than that in HCs (1.56 ± 0.33 vs 2.07 ± 0.53, P < 0.0001). The mean expression of log10 LINC00472 in the advanced stage was higher than those in the early stage and HCs (2.99 ± 0.87 vs 1.81 ± 0.83, P < 0.0001; 2.99 ± 0.87 vs 1.77 ± 0.84, P < 0.0001, respectively). The mean expression of LINC01257 in both the early stage and advanced stage were up-regulated compared with HCs (3.88 ± 1.55 vs 2.37 ± 0.96, P < 0.0001; 3.57 ± 1.79 vs 2.37 ± 0.96, P < 0.0001, respectively). The areas under the curves (AUC) for S100A6, LINC00312, log10 LINC00472 and LINC01257 in PBC diagnosis were 0.759, 0.7292, 0.6942 and 0.7158, respectively. Furthermore, the AUC for these four genes in PBC staging were 0.666, 0.661, 0.839 and 0.5549, respectively. The expression levels of S100A6 mRNA, log10 LINC00472, and LINC01257 in plasma of PBC patients were decreased (2.35 ± 1.02 vs 3.06 ± 1.04, P = 0.0018; 1.99 ± 0.83 vs 2.33 ± 0.96, P = 0.036; 2.84 ± 0.92 vs 3.69 ± 1.54, P = 0.0006), and the expression level of LINC00312 was increased (1.95 ± 0.35 vs 1.73 ± 0.32, P = 0.0007) after treatment compared with before treatment using the paired t-test. Relative expression of S100A6 mRNA was positively correlated with log10 LINC00472 (r = 0.683, P < 0.0001); serum level of collagen type IV was positively correlated with the relative expression of log10 LINC00472 (r = 0.482, P < 0.0001); relative expression of S100A6 mRNA was positively correlated with the serum level of collagen type IV (r = 0.732, P < 0.0001). The AUC for the four biomarkers obtained in the validation set were close to the training set.CONCLUSIONThese four genes may potentially act as novel biomarkers for the diagnosis of PBC. Moreover, LINC00472 acts as a potential biomarker for staging in PBC.

Secretin Treatment Promotes Hepatic Progenitor Cell Activation, Ductal‐Canalicular Junction Formation and Amelioration of Liver Damage in a Model of Late‐Stage Primary Biliary Cholangitis

Introduction PBC is a cholestatic disease characterized by bile duct loss, ductular reaction and liver fibrosis. The canals of Hering (CoH) are the site of ductulo-canalicular junction (DCJ) that maintain proper bile drainage from the liver, and contain the hepatic progenitor cell (HPC) population. CoH loss is noted in PBC and correlates with scoring. Secretin binds to its receptor (SR, expressed by large cholangiocytes) to promote biliary proliferation during cholestasis; however, during large duct damage, small cholangiocytes acquire large cholangiocyte phenotypes including SR expression. Secretin/SR expression is downregulated in late-stage PBC; therefore, we aimed to understand the role of the secretin/SR axis during PBC. We hypothesized that secretin treatment restores DCJ through HPC activation and ameliorates liver phenotypes associated with late-stage PBC. Methods Female 34 wk old wild-type (WT) and dominant-negative transforming growth factor-β receptor II (dnTGFβRII, model of late-stage PBC) mice were treated with control or secretin (2.5 nmol/kg BW) for 8 wk by IP implanted minipumps. Cholangitis activity, hepatitis activity and bile duct loss were determined by the Nakanuma scoring system. Ductular reaction was measured by semi-quantitative CK-19 immunostaining. Liver fibrosis was evaluated by Sirius Red staining, collagen-1a immunostaining and qPCR for α-SMA. Hepatic stellate cell (HSC) presence was determined by desmin immunostaining. Cholestasis was indicated by hepatic total bile acid (TBA) levels using EIA. DCJ presence was evaluated by co-immunostaining for ABC transporter bile salt export pump (BSEP)/CK-19 and semi-quantification. HPC activation was assessed by Sox9 staining and semi-quantification. Results dnTGFβRII mice had an increase in cholangitis and hepatitis activity; however, 8 wks of secretin treatment significantly reduces cholangitis, but not hepatitis, activity. dnTGFβRII mice present with ductular reaction and significant bile duct loss, which was reversed with 8 wks secretin treatment. Hepatic TBA levels, liver fibrosis and HSC presence are increased in dnTGFβRII mice, but significantly reduced following 8 wks of secretin treatment. In line with human studies, dnTGFβRII mice have significant loss of DCJ and reduced HPC number; however, 8 wks of secretin treatment restores these parameters. Conclusion Long-term secretin treatment (i) reduces ductular reaction, (ii) reverses bile duct loss, (iii) decreases liver fibrosis, (iv) promotes HPC activity and (v) restores DCJ presence in a model of late-stage PBC. Secretin maintenance of the DCJ may be due to (i) the restoration of bile flow and release of toxic bile acids or (ii) secretin binding to SR acquired on small cholangiocytes/HPCs. Treatment with secretin may be a therapeutic option for late-stage PBC patients.

Type IV collagen as a potential biomarker of metastatic breast cancer

No reliable, non-invasive biomarker of metastatic breast cancer (mBC) exists: circulating CA15-3 (cCA15-3) is the marker mostly used to monitor mBC. Circulating collagen IV (cCOLIV) has been evaluated in other metastatic cancers and has been found to be a promising biomarker. The overarching aim of this study was to evaluate cCOLIV as a potential biomarker in patients with mBC. The first aim was to determine the levels of cCOL IV and cCA15-3 in patients with healthy controls, primary breast cancer (pBC) and mBC. The second aim was to compare levels of cCOLIV and cCA15-3 in patients with different metastatic sites of BC. The third aim was to investigate the prognostic value of cCOLIV and cCA15-3 for mBC patients. The fourth aim was to analyse whether a combination of the two biomarkers was more accurate in detecting mBC than a single marker. Lastly, we investigated the tissue expression levels of COLIV in BC bone metastases (BM) and liver metastases (LM). Plasma levels of cCOLIV and cCA15-3 from healthy controls and patients with pBC and mBC were measured. COLIV expression in tissue from patients with LM and BM was analysed using immunohistochemistry. Clinical and survival data were collected from medical charts. The levels of cCOLIV and cCA15-3 were significantly elevated in mBC patients compared with healthy controls and pBC patients. No differences in cCOLIV and cCA15-3 levels were found based on the metastatic site. High levels of cCOLIV, but not cCA15-3, correlated with poorer survival. cCOLIV alone and the combination of cCA15-3 and cCOLIV were superior to cCA15-3 at detecting mBC. COL IV was highly expressed in the tissue of LM and BM. Our study suggests that cCOLIV is a potential marker to monitor patients with BC.

Abstract PS9-57: Estimation of willingness-to-pay for breast cancer treatments through contingent valuation method in Japanese breast cancer patients (JCOG1709A); preliminary study findings

Abstract BackgroundIn 2019, the Japanese government has decided to introduce the economic valuation to healthcare system. However, the threshold for determining cost-effectiveness-analysis has yet to be adequately studied. Therefore, we have designed a study to examine the financial value of “life” and “health” based on willingness-to-pay (WTP) considered by approximately 1,800 patients with primary and metastatic breast cancer (PBC and MBC) in Japan. Preliminary study has been conducted to set the amount of WTP prior to the main study. Methods This study is conducted by six leading hospitals under the Breast Cancer Study Group of Japan Clinical Oncology Group (JCOG). 168 patients (84 patients with PBC and 84 patients with MBC) from 20 to 79 years of age paying medical costs were examined their WTP for setting up the dichotomous-choice method survey form. Virtual scenario-specific treatments to avoid the recurrence and death of breast cancer for one year for PBC patients, and to prolong the survival period for one year for the patients with MBC were presented. The patients were evaluated how much money would pay to receive the treatment in a self-written answer. The amount of WTP that will be presented in the main study was determined by calculating the median price from these responses based on the protocol. In addition, we conducted surveys of quality-of-life by EQ-5D-5L, social background on patients, and of breast cancer medical condition on doctors. Results Completed surveys were collected from 166 doctors (98.8%) and 139 patients (82.7%). The range of the amount of WTP for scenario-specific treatments among the patients with the patients with PBC was from 0 to 5 million Japanese yen (JPY) (50,000 USD), and among the patients with MBC was from 0 to 10 million JPY (100,000 USD). In accordance with the criteria set forth in advance in the protocol, the amount of WTP that will be presented in the main study was set at 5 million JPY in both PBC and MBC patients. Conclusion We have fixed the cost that will be presented in the main study based on the findings of our preliminary study. The main study will commence around September 2020. Citation Format: Tsuguo Iwatani, Fumikata Hara, Tadahiko Shien, Masato Takahashi, Norikazu Masuda, Hiroyuki Yasojima, Yasuaki Sagara, Tomonori Mizutani, Keita Sasaki, Kenichi Nakamura, Haruhiko Fukuda, Takaru Shiroiwa, Hiroji Iwata. Estimation of willingness-to-pay for breast cancer treatments through contingent valuation method in Japanese breast cancer patients (JCOG1709A); preliminary study findings [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS9-57.

An enhanced level of LAMP-2A participates in CD4+T cell hyperactivity in patients with primary biliary cholangitis.

BackgroundPrimary biliary cholangitis (PBC) is an immune-mediated chronic cholestasis, in which T cell homeostasis plays an important role. Lysosomal-associated membrane protein 2 isoform A (LAMP-2A) has been implicated in the regulation of CD4+T cell responses.MethodsWe comprehensively evaluated the immunobiology of CD4+T cells in patients with PBC (PBC, n=42), chronic hepatitis B (CHB, n=20), and healthy control subjects (HC, n=20) by flow cytometry including activation status and LAMP-2A expression. Additionally, we investigated the activation responses of PBC-naïve CD4+T cells by stimulation in vitro and tested the changes caused by deleting the gene encoding LAMP-2A.ResultsFirstly, we found an increased activation status of circulating CD4+T cells from PBC patients compared to the HC subjects, and PBC-naïve CD4+T cells showed enhanced responses after stimulation in vitro. Secondly, PBC-naïve CD4+T cells expressed a significantly higher level of LAMP-2A compared to the HC and CHB groups [PBC vs. HC, 1,954.74 (1,254.28–3,057.14) vs. 1,542.12 (961.18–2,277.98), P=0.03; vs. CHB, 1,153.59 (726.87–1,275.48), P=0.02], and the overreactions of PBC-naïve CD4+T cells could be reversed by interfering with LAMP-2A expression in vitro. Thirdly, the LAMP-2A expression level of PBC-naïve CD4+T cells was related to disease severity and drug response.ConclusionsAn abnormally increased LAMP-2A expression of PBC-naïve CD4+T cells might be related to excessive activation responses. LAMP-2A could be a novel therapeutic target for the treatment of PBC by reversing excessive responses and consequently reducing biliary injury.

Decreased infiltration of CD4+ Th1 cells indicates a good response to ursodeoxycholic acid (UDCA) in primary biliary cholangitis

BackgroundPrimary biliary cholangitis (PBC) is characterized by nonsuppurative destructive cholangitis and is thought to be an autoimmune disorder. Currently, ursodeoxycholic acid (UDCA) is the only FDA approved first-line therapy for PBC, but up to nearly one-third of patients do not achieve a complete response to this treatment. Adaptive immune cells, including T cells and B cells, have been found in the portal tracts and the bile duct epithelium and play a role in the pathogenesis of PBC, but the importance of these cells for evaluating the therapeutic response to UDCA in PBC has not yet been studied. MethodsIn this study, we collected liver puncture biopsy specimens from 34 matched patients with PBC before and after UDCA treatment and investigated the relationship between the infiltration of adaptive immune cells and the treatment response to UDCA. The extent of immune cell infiltration was determined by immunohistochemical analysis. Responses were defined based on Paris-I criteria. ResultsAfter 1 year of treatment, 25/34 patients responded to UDCA treatment according to Paris-I criteria (responders), and 9/34 patients were nonresponders. Immunohistochemical analysis showed that UDCA responders exhibited significantly less CD4+ T cell infiltration after UDCA treatment than before (50.4 ± 7.5/HPF vs 30.0 ± 7.9/HPF, P = 0.002). In contrast, UDCA nonresponders exhibited significantly more CD4+ T cell infiltration after UDCA treatment than before (32.2 ± 8.0/HPF vs 75.0 ± 13.9/HPF, P = 0.045). Moreover, patients who exhibited a reduction in CD4+ T cell infiltration after UDCA treatment had a higher response rate than those that exhibited an increase in CD4+ T cell infiltration (85.7 % vs 53.8 %, P = 0.041). However, CD3+ T cell, CD8+ T cell, and CD20+ B cell infiltration was not significantly different before and after treatment in either UDCA responders or nonresponders. Furthermore, we found that the number of infiltrating T-bet+ Th1 cells was much lower after UDCA treatment than before in responders (10.5 ± 5.7/HPF vs. 5.16 ± 4.0/HPF, P = 0.0214) but much higher in nonresponders after treatment than before (1.89±1.2/HPF vs. 12.3±5.4/HPF, P = 0.043). However, there was no difference in the extent of GATA3+ Th2 or FOXP3+ Treg infiltration before and after treatment in either UDCA responders or nonresponders. ConclusionCollectively, our results suggest that a decrease in the number of liver-infiltrating CD4+ Th1 cells is associated with a good response of PBC patients to UDCA treatment. Immunohistochemical analysis of CD4 and T-bet in PBC liver specimens may be a potential approach for evaluating the therapeutic response to UDCA.

Monocytes as Potential Mediators of Pathogen-Induced T-Helper 17 Differentiation in Patients With Primary Sclerosing Cholangitis (PSC).

T cells from patients with primary sclerosing cholangitis (PSC) show a prominent interleukin (IL)-17 response upon stimulation with bacteria or fungi, yet the reasons for this dominant T-helper 17 (Th17) response in PSC are not clear. Here, we analyzed the potential role of monocytes in microbial recognition and in skewing the T-cell response toward Th17. Monocytes and T cells from blood and livers of PSC patients and controls were analyzed ex vivo and in vitro using transwell experiments with cholangiocytes. Cytokine production was measured using flow cytometry, enzyme-linked immunosorbent assay, RNA in situ hybridization, and quantitative real-time PCR. Genetic polymorphisms were obtained from ImmunoChip analysis. Following ex vivo stimulation with phorbol myristate acetate/ionomycin, PSC patients showed significantly increased numbers of IL-17A-producing peripheral blood CD4+ T cells compared to PBC patients and healthy controls, indicating increased Th17 differentiation in vivo. Upon stimulation with microbes, monocytes from PSC patients produced significantly more IL-1β and IL-6, cytokines known to drive Th17 cell differentiation. Moreover, microbe-activated monocytes induced the secretion of Th17 and monocyte-recruiting chemokines chemokine (C-C motif) ligand (CCL)-20 and CCL-2 in human primary cholangiocytes. In livers of patients with PSC cirrhosis, CD14hiCD16int and CD14loCD16hi monocytes/macrophages were increased compared to alcoholic cirrhosis, and monocytes were found to be located around bile ducts. PSC patients show increased Th17 differentiation already in vivo. Microbe-stimulated monocytes drive Th17 differentiation in vitro and induce cholangiocytes to produce chemokines mediating recruitment of Th17 cells and more monocytes into portal tracts. Taken together, these results point to a pathogenic role of monocytes in patients with PSC.

Seronegative Primary Biliary Cholangitis: Epidemiological, Clinical, Therapeutic and Progressive Features

Introduction: Seronegative PBC (specific AMA antibodies are negative) is a particular clinical form of PBC.The aim of this study was to determine the clinical-biological and evolutionary features of seronegative PBCs. Methods: The medical data of 42 PBC patients were evaluated and the AMA positive and negative groups were compared. Results: 42 cases of PBC were collected. These were 41 women and 1 man. The mean age at diagnosis was 46 years (25-70 years). AMA Abs were most often positive (n = 34). Other antibodies were found in 22 patients. In our series, 9.75% of cases had seronegative PBC. By comparing the 2 groups of PBC with or without AMA Abs, depending on age, sex, degree of fibrosis and response to AUDC, no statistically significant difference was noted. Conclusion: In our series, the prevalence of seronegative PBC was 9.75%. Its clinical-biological and evolutionary profile under treatment does not seem to differ from that of seropositive PBC.

The role of factor Y in primary biliary cirrhosis

Abstract Primary biliary cirrhosis (PBC) is an autoimmune disease which is characterized by bile duct injuries result from excessive immune responses. In female over the age of 40, the incidence of PBC has been estimated to be one out of 1000 people. Liver transplantation is required for patients who progress to end-stage PBC. However, risk factors and causes of PBC remain elusive. We performed whole-exome sequencing in a family with four sisters diagnosed with PBC and one healthy half-sister. Additionally, Sanger sequencing was conducted with 62 unrelated PBC patient specimens. Here, we identified a potential PBC causative gene factor Y, a regulator of cell death and inflammation. We investigated whether factor Y plays a role in PBC using factor Y null human macrophage cell line generated by CRIPSR/Cas9 technology. We found ablation of factor Y induces inflammatory cytokines compared with control cells upon treatment with LPS or TNF. Surprisingly, molecules secreted by factor Y null cell line induced fibrosis in human hepatic stellate cell, LX-2. These results indicate that factor Y is a potential target for treatment of PBC.

Profiling of the pattern of the human TRB/IGH–CDR3 repertoire in primary biliary cholangitis patients

IntroductionPrimary biliary cholangitis (PBC) is characterized by lymphocyte cell-induced immune destruction of cholangiole. However, the immunological characteristics of peripheral blood cells in PBC patients remain unknown. This study was designed to reveal the differences in the immunological characteristics between PBC patients and healthy adults. MethodsWe performed high-throughput sequencing to determine the TRB–CDR3 and IGH–CDR3 repertoires of T and B cells in 19 healthy controls and 29 PBC patients. Different immunological characteristics, such as distinctive complementarity determining region 3 (TRB–CDR3) lengths, usage bias of V and J segments, and random nucleotide addition were identified in PBC and healthy control (HC) groups. ResultsThe diversity of TRB–CDR3 was significantly lower in the PBC group compared with the HC group. CDR3 and the N addition length distribution were significantly changed compared with the HC group. It appeared that the PBC group had more short N additions and the HC group had more long N additions in the TRB–CDR3 repertoire. The results also revealed a set of PBC-associated clonotypes compared with the HC group. ConclusionThis study suggested that PBC is a complex autoimmune disease process with evidence of different TRB–CDR3 rearrangements compared with healthy adults that share IGH–CDR3 peptides with some autoimmune diseases. This new insight may contribute to a better understanding of the immune functions of PBC patients and benefit efficient applications of PBC diagnosis and treatments.