Taipei Institute of Pathology

A biowaiver generally refers to the request to waive an in vivo bioequivalence study. A biowaiver may be granted not only based on the Biopharmaceutics Classifications System (BCS) but also for many immediate-release dosage forms based on pre-defined criteria. The current paper summarises the results from a survey of the biowaiver requirements for cutaneous/topical products (topical solutions, gels, suspensions, ointments, and creams), ear/otic and ophthalmic solutions and suspensions, enemas in solution and suspension, and vaginal solid dosage forms and suppositories defined by the participants of the Bioequivalence Working Group for Generics (BEWGG) of the International Pharmaceutical Regulators Programme (IPRP). A review of the results from the survey indicates that there is a trend towards convergence when the dosage forms are less complex; however, the most common approach used by each of the participants was a case-by-case approach given that most participants do not have well-defined guidelines to support all possible scenarios. Notwithstanding the differences, disseminating information is the first step towards regulatory convergence regarding biowaivers for certain dosage forms and will be useful for pharmaceutical companies currently developing generic medicinal products for countries represented by IPRP participants.

Hepatocellular carcinoma (HCC) progression is tightly linked to metabolic reprogramming and immune evasion. However, the transcriptional networks driving these processes remain misunderstood. Here, we identified a novel regulatory axis wherein the transcription factor SOX4 formed a stress-responsive complex with NRF2, as confirmed by co-immunoprecipitation and proximity ligation assay. This process was orchestrated via p62-mediated disruption of the KEAP1-SOX4 complex. The SOX4-NRF2 complex directly activated Phosphoserine Phosphatase (PSPH) transcription-as revealed by luciferase reporter and chromatin immunoprecipitation-enhancing serine biosynthesis and downstream metabolites critical for oxidative phosphorylation (OXPHOS) and redox balance. Inhibition of SOX4 or NRF2 impaired PSPH expression, exacerbated oxidative damage-marked by elevated 4-hydroxynonenal-and increased sensitivity to sorafenib treatment in HCC cells. Furthermore, PSPH-driven metabolites, particularly serine, fostered M2-like macrophage polarization, thereby potentially contributing to the promotion of an immunosuppressive tumor microenvironment. Analysis of HCC specimens from TCGA and clinical cohorts confirmed that high SOX4/NRF2/PSPH expression was correlated with increasing M2 macrophage infiltration and poor patient prognosis. Our findings revealed a previously unrecognized SOX4-NRF2-PSPH regulatory loop that coupled cancer metabolism with immune modulation. Targeting this axis may offer a promising therapeutic avenue to simultaneously disrupt metabolic support and immune evasion in HCC.

Background:Breast cancers (BRCs) can be classified into 6 molecular subtypes based on gene expression profiles. Previous research suggests that tumor-infiltrating lymphocytes are associated with metastasis-free survival (MFS) in triple-negative and HER2-overexpressing BRC.Objectives:Our study aims to investigate further how the immune response (IR) may impact MFS in different molecular subtypes of BRC.Design:A single hospital-based retrospective cohort study.Methods:A training series of 327 BRCs was used to identify 297 IR transcripts that were correlated with the T cell–associated CD3D transcript or the B cell-associated CD19 transcript. Using these IR transcripts, each of the 6 molecular subtypes was hierarchically clustered into high and low immune responders. An IR score based on the average of the 297 IR transcripts was determined for each BRC. Correlations between the IR score and 3 signatures for IR or response to immune checkpoint inhibition therapy (ICIT) were investigated. A series of 884 BRCs from public datasets was used for confirmation, and the other independent series of 988 BRCs was used for validation.Results:For subtype I, high immune responders had a statistically significantly better MFS than low immune responders in all the training, confirmation, and validation series by Kaplan-Meier survival analysis (P = .0039, .049, .039, log-rank test). The same trend was observed for subtype II (P = .16, .052, .015) and subtype IV (P = .0078, .0002, .12). Our IR scores were linearly correlated with the Teschendorff, the T-effector and IFNg, and the T–cell inflamed signatures for IR or ICIT. The IR scores were also linearly correlated with the expression of 6 different immune checkpoint genes.Conclusions:Tumor IR is a biomarker for MFS for BRCs of I, II, and IV subtypes. Our study supports the potential use of the IR score for identifying patients responsive to ICIT.

The purpose of this study was to investigate aqueous humor total antioxidant capacity (TAC) and ascorbic acid (AA) levels of patients with diabetic retinopathy (DR), and their correlation with serum glycated hemoglobin (HbA1c) levels. This case-control study enrolled 177 eyes from 177 patients that underwent cataract surgery between April 2019 and November 2022 in Chang Gung Memorial Hospital. Group 1 comprised 103 eyes from patients without diabetes; group 2 comprised 40 eyes from patients with diabetes but without DR; and group 3 comprised 34 eyes from patients with DR. Aqueous humor samples were collected and analyzed for TAC and AA levels using the ferric reducing/antioxidant AA assay. Aqueous humor TAC and AA levels were decreased in group 3 compared with group 2 and group 1 (P < 0.001 and P < 0.001). Aqueous humor TAC and AA levels were independent protective factors against DR in the diabetes population, with adjusted odds ratios of 0.109 (95% confidence interval [CI] = 0.017-0.719, P = 0.021) and 0.095 (95% CI = 0.012-0.726, P = 0.023), respectively. Aqueous humor TAC and AA concentrations were weakly negatively correlated with serum HbA1c levels, with Pearson's correlation coefficients of -0.339 (P = 0.013) and -0.341 (P = 0.012), respectively. TAC and AA had protective effects against DR. Decreased aqueous humor TAC and AA levels may serve as potential biomarkers for DR among the diabetes population. This study highlights the role of antioxidants in DR and provides foundation for future research on AA supplementation's potential for the prophylaxis and treatment of DR.

e16460 Background: Pancreatic cancer (PC) is one of the deadliest malignancies, largely due to the lack of effective early detection methods. Traditional biomarkers like CA19-9 and CEA have limited sensitivity and specificity, particularly in early-stage cases. The EG-Pancreatic Blood Test-E1, a novel assay targeting circulating cfDNA methylation of ZFP30 , offers a promising alternative for early PC detection. This study evaluates its diagnostic performance in Western and Asian populations, with a focus on early-stage detection. Methods: Plasma samples were collected from early-stage PC patients and healthy controls across Western and Asian cohorts. The EG-Pancreatic Blood Test-E1 utilizes a multi-step process, including cfDNA extraction, bisulfite conversion, and methylation-specific quantitative PCR (qPCR) to quantify ZFP30 methylation levels. Sensitivity, specificity, and overall accuracy were evaluated and compared against conventional biomarkers, CA19-9 and CEA. The diagnostic efficacy of ZFP30 methylation was assessed using receiver operating characteristic (ROC) curve analysis and area under the curve (AUC) measurements. Results: The EG-Pancreatic Blood Test-E1 demonstrated exceptional diagnostic performance in detecting pancreatic cancer, particularly in early-stage patients. In the Western cohort, the assay achieved a sensitivity of 92.3%, a specificity of 96.2%, and an overall accuracy of 93.2%. Receiver operating characteristic (ROC) curve analysis confirmed its superior diagnostic capability, with an area under the curve (AUC) of 0.95, surpassing the performance of conventional biomarkers CA19-9 and CEA. When evaluated across both Western and Asian populations, the test exhibited a remarkable sensitivity of 96.0%, specificity of 92.4%, and overall accuracy of 92.5%. The consistency of ZFP30 methylation levels across diverse patient subgroups underscores its robustness as a biomarker and its potential for widespread clinical application. Conclusions: The EG-Pancreatic Blood Test-E1, utilizing ZFP30 cfDNA methylation, represents a groundbreaking advancement in pancreatic cancer diagnostics. Its exceptional accuracy, non-invasive nature, and superior performance over conventional markers position it as a transformative tool for early detection across diverse populations. This innovation has the potential to redefine precision oncology and significantly improve patient survival rates by enabling earlier diagnosis and timely treatment.

In this study, we aim to evaluate the risk factors of myopia undercorrection in recipients of second-generation keratorefractive lenticule extraction (KLEx) surgery. A retrospective case-control study was performed, and patients who received second-generation KLEx surgery were enrolled. The cases with myopia undercorrection were matched to non-myopia undercorrection cases with a 1:4 ratio according to age, and a total of 22 and 88 eyes were categorized into the undercorrection and control groups, respectively. Demographic, refractive, topographic, and surgical data were collected preoperatively. A generalized linear model was operated to evaluate the potential risk factors for myopia undercorrection. The uncorrected distance visual acuity (UDVA) at three months postoperation was significantly better in the control group (p = 0.006), and residual myopia and SE were significantly higher in the undercorrection group during the whole follow-up period (all p < 0.001). The UDVA value showed a trend of improvement in the control group (p < 0.001), and the changes to SE and residual myopia were significantly lower in the control group (both p < 0.001). Regarding the risk factors for myopia undercorrection in the whole population and the high-myopia population, a higher manifest sphere power, higher steep keratometry (K), higher topographic cylinder, lower central corneal thickness (CCT) at apex, higher CCT difference and lower residual stromal thickness (RST) correlated to myopia undercorrection (all p < 0.05). In the low-myopia population, only higher myopia and lower RST correlated to myopia undercorrection (both p < 0.05). In conclusion, a high-sphere power and irregular topographic pattern correlated to myopia undercorrection after the second KLEx surgery, especially for individuals with high myopia.

Abstract A 61-year-old woman presented with a 7-month history of a subcutaneous nodule in her left axilla. On examination, a 1.5 × 1.5-cm mobile rubbery nodule was noted in her left axilla. Biopsy of the lesion revealed neoplastic cuboidal epithel­ial cells in a microcystic and papillary-cystic pattern adjacent to non-neoplastic accessory breast tissue in the deep subcutaneous layer. The cells had monotonous nuclei, eosinophilic cytoplasm and distinct nucleoli. Immunohistochemically, the tumour cells were diffusely positive for S100, CK5/7, GATA3 and pan-TRK, while oestrogen receptor, progesterone receptor and human epidermal growth factor receptor 2 (HER2) stained negative. Further image investigation revealed no evidence of malignancy in the breasts, lymph nodes or salivary glands. A diagnosis of secretory carcinoma (SC) arising from the accessory breast was made based on the clinicopathological correlation. The main differential diagnoses included primary cutaneous SC (PCSC), metastatic breast SC, and metastatic salivary SC. In this case, the presence of accessory breast tissue adjacent to the neoplasm, tumour involvement in the deep dermal–subcutaneous layer, and negative primary tumour in the breast and salivary glands favoured SC of the accessory breast. A wide excision with 1-cm safety margin was performed. The next-generation sequencing analysis confirmed presence of neurotrophic tyrosine receptor kinase 3 (NTRK3) fusion genes. SC of the breast is a very rare subtype of breast carcinoma, accounting for only 0.15% of all breast cancers. Clinically, it manifests as an indolent subcutaneous mass on the breast. Pathologically, SC is characterized by tumour cells exhibiting intracellular and extracellular secretory materials, forming patterns including cribriform, papillary-cystic, lobulated and microcystic. Immunohistochemically, SC expresses S100, basal-like markers (such as CK5/6), epidermal growth factor receptor and pan-TRK, and is usually negative for oestrogen receptor, progesterone receptor and HER2. Studies have shown that these carcinomas develop due to t(12,15) ETV6–NTRK3 gene translocation. Notably, the genetic alteration is not exclusive to a specific origin site, emphasizing the importance of clinicopathological correlation for accurate diagnosis. Furthermore, while most PCSCs are superficially located in the dermis, SC originating from accessory breast is found in the deeper subcutaneous layer, often adjacent to breast tissue. To our knowledge, this is the third report of SC arising from the axillary accessory breast. Our patient highlights the significance of considering SC of the accessory breast as a potential differential diagnosis for indolent axillary masses.

Abstract The 2021 WHO classification underscores the importance of molecular data integration in Central Nervous System (CNS) tumor diagnostics. However, currently used assays have disadvantages due to technical complexity, required equipment and reagent cost, as well as lengthy turnaround times. In response to these challenges, we introduce Rapid-CNS2 and MNP-Flex. Rapid-CNS2, an adaptive sampling-based nanopore sequencing workflow, offers real-time methylation classification and DNA copy-number information within a 30-minute window, suitable for intra-operative settings, followed by comprehensive molecular profiling within 24 h, covering the complete spectrum of diagnostically and therapeutically relevant information for the respective entity. We have prospectively validated Rapid-CNS2 in a multi-center setting on 223 samples. For even more widespread use of methylation-based CNS tumor classification, we developed MNP-Flex, a platform-agnostic methylation classifier encompassing 184 CNS tumor classes. MNP-flex achieved 92% accuracy across a global validation cohort of 78,000 samples spanning five different technologies. These innovations represent a significant advancement in CNS tumor diagnostics, making rapid, actionable molecular insights more widely and more rapidly available, which is crucial for personalized treatment strategies. Their integration streamlines the diagnostic process, broadening access to accurate molecular classification and promising improved patient outcomes in neurooncology on a global scale. MNP-Flex is available as a web-service https://mnp-flex.org and the Rapid-CNS2 workflow is available on Github.