Immunohistochemical Staining Intensity Research Articles

Expression of indoleamine-2,3-dioxygenase 1 in sebaceous glands and related tumors of the eyelid: a potential diagnostic target

ABSTRACT Purpose Mechanisms that modulate the immune response hold great potential as viable strategies for combating cancer. Among these, novel molecular compounds that inhibit IDO1 have emerged as one of the most promising therapeutic opportunities for impeding tumor growth. The present objective is to describe the expression of indoleamine-2,3-dioxygenase 1 (IDO1) in sebaceous glands and study its role in diagnosing sebaceous carcinoma of the eyelid. Methods We conducted a clinical-pathologic study of IDO1 expression in healthy sebaceous glands, sebaceous adenomas (SA), sebaceous hyperplasia (SH), and sebaceous carcinoma (SC). A comparison of immunohistochemical staining intensity and distribution for IDO1 was performed between all groups. Clinical data was correlated with IDO1 staining. Results The study included 38 patients: 6 controls, 5 SA, 6 SH, and 18 SC. The mean H-scores of the control group, SA, SH, and SC were 0, 110, 127.5, and 201.9, respectively. There was a significant relationship between IDO1 h-scores and both patient age and tumor stage (p-values <0.001). There was a significant increase in IDO1 expression between SC and all other groups. Conclusions Our study demonstrated a statistically significant upregulation and reactivity of IDO1 in sebaceous gland tumors. Prior to these findings, clinical management centered around surgical excision. The use of molecular compounds such as indoximob, epacadostat, BMS-986205 and navoximod, that directly target IDO1 opens new possibilities for targeting these tumors, improving clinical outcomes, and minimizing the morbidities often associated with surgery.

Imaging with [ 89 Zr]Zr-DFO-SC16.56 anti-DLL3 antibody in patients with neuroendocrine tumors of the prostate: A phase 1/2, first-in-human trial.

e15057 Background: Delta-like ligand 3 (DLL3) is expressed on the tumor cell surface of some neuroendocrine prostate cancer (NEPC). We assessed the safety and feasibility of the DLL3-targeted imaging tracer [ 89 Zr]Zr-DFO-SC16.56, which is composed of the anti-DLL3 antibody SC16.56 conjugated to p-SCN-Bn-deferoxamine (DFO) and zirconium-89 in patients with NEPC. Methods: We conducted an open-label, first-in-human study of immunoPET/CT imaging with [ 89 Zr]Zr-DFO-SC16.56. The initial phase I included patients with small-cell lung cancer (n=3) that showed high tumoral uptake of the diagnostic tracer without any major adverse events. The expansion cohort included NEPC patients that received a single infusion of [ 89 Zr]Zr-DFO-SC16.56 at the same activity (1-2 mCi) and mass dose (2-3 mg) as in the initial cohort followed by a single PET-CT scan 3–5 days later. Retrospectively collected tumor biopsy samples were assessed for DLL3 by immunohistochemistry (IHC). DLL3 positive expression was defined as ≥ 5% weak (1+) IHC staining intensity. The primary endpoint of the expansion cohort was to determine the correlation between tumor uptake of [ 89 Zr]Zr-DFO-SC16.56 with expression of DLL3 as determined by IHC. Results: Between April 2023, and October 2024, 11 men with NEPC were enrolled, with a median age of 60 years (range 51–80). A single immunoPET/CT scan on day 3–5 post-administration could delineate DLL3-avid tumors in 6 (55%) of 11 patients. A range in tumoral uptake was observed within individual patients as well as the entire cohort, with a wide range in maximum standardized uptake value (from 0·4 to 75·5). Tumoral uptake by [ 89 Zr]Zr-DFO-SC16.56 was associated with protein expressionin 8 (80%) of 10 patients who had DLL3 IHC performed.None of the patients had any adverse events noted, and no clinically meaningful changes were observed in vital signs or laboratory parameters post injection. Conclusions: DLL3 PETCT imaging of patients with NEPC is safe and feasible. These results show the potential utility of [ 89 Zr]Zr-DFO-SC16.56 for non-invasive in vivo detection of NEPC patients. Clinical trial information: 19-292 . Characteristics of patients included in the study. Age DLL3 avid SUV max of highest DLL3-avid lesion (anatomic location of most avid lesion) Blood uptake from day 3–5 images, SUV mean Biopsy location (immunohistochemistry H-score) Patient 1 71 No 2·2 (liver) 3·7 Liver (0) Patient 2 58 Yes 75·5 (liver) 2·4 Liver (220) Patient 3 57 No 0·4 (Sacrum) 0·7 Prostate (0) Patient 4 60 Yes 11·3 (Humerus) 1·3 Liver (190) Patient 5 80 Yes 39·9 (liver) 5·8 Pleurae (280) Patient 6 62 Yes 10·4 (Sacrum) 7·1 Liver (0) Patient 7 51 No 6·7 (lymph node) 6·5 Lymph node (170) Patient 8 60 Yes 36·4 (liver) 6·3 Liver (230) Patient 9 55 Yes 20·3 (liver) 7·0 R Liver (170) Patient 10 74 No 3·9 (vertebrae) 7·5 Liver (NA) Patient 11 65 No 1·7 (vertebrae) 5·4 Lymph node (0) NA=not applicable.

A multiregional, randomized, controlled, open-label, phase 3 study of the anti-claudin18.2 (CLDN18.2) antibody-drug conjugate (ADC) arcotatug tavatecan (IBI343) in gastric or gastroesophageal junction adenocarcinoma (G/GEJA): Trial in progress.

TPS4201 Background: CLDN18.2 has been a validated therapeutic target for G/GEJA. As a next-generation ADC, arcotatug tavatecan (IBI343) composed of anti-CLDN18.2 monoclonal antibody conjugated to exatecan (topoisomerase I inhibitor) with unique IgG1 Fc silencing to attenuate antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. Previous phase 1 studies of IBI343 observed manageable safety profiles with encouraging efficacy in G/GEJA, pancreatic ductal adenocarcinoma and biliary tract cancer (2024 ASCO Annual Meeting [3037], ESMO GI 2024 [396MO], ESMO Asia 2024 [132MO]). Here, we present the trial in progress of a phase 3 study (G-HOPE-001, NCT06238843) evaluating efficacy and safety of IBI343 monotherapy versus treatment of investigator’s choice in previously treated patients (pts) with CLDN18.2-positive G/GEJA. Methods: This multiregional, randomized, controlled, open-label, phase 3 study planned to enroll 450 pts. Main inclusion criteria are: 1) locally advanced unresectable or metastatic G/GEJA; 2) positive CLDN18.2, defined as immunohistochemical (IHC) membrane staining intensity ≥ 2+ in ≥ 75% of tumor cells as measured by the VENTANA CLDN18 (43-14A) Assay; 3) radiologically evaluable disease, measurable and/or non-measurable disease per RECIST v1.1; 4) received and progressed on 2-4 prior regimens of systemic therapy which must include a fluoropyrimidine, platinum, and a taxane or irinotecan. Main exclusion criteria are: 1) positive HER-2, defined as IHC 3+ or IHC 2+/in situ hybridization+; 2) history of treatment with topoisomerase inhibitor-based ADCs. Pts are randomized in a 1:1 ratio to receive IBI343 6mg/kg Q3W in the experimental arm or to receive treatment of investigator’s choice including irinotecan, paclitaxel, or trifluridine/tipiracil in the control arm. Stratification factors include region (Asian country/region other than Japan vs. European Union and United States vs. Japan), primary site of the tumor (stomach vs. gastroesophageal junction) and history of prior gastrectomy (yes vs. no). The primary endpoints are progression-free survival (PFS) per RECIST v1.1 and overall survival (OS). The secondary endpoints include objective response rate (ORR), disease control rate (DCR), duration of response (DoR) and time to response (TTR) per RECIST v1.1, quality of life (QoL), safety, pharmacokinetics (PK) and immunogenicity. The trial is currently enrolling pts in China and Japan. Clinical trial information: NCT06238843 .

Astaxanthin alleviates fipronil-induced neuronal damages in male rats through modulating oxidative stress, apoptosis, and inflammatory markers

Fipronil (FPN) is an effective pesticide for veterinary and agricultural use; however, it can induce neurotoxic effects on non-target organisms after accidental exposure. Astaxanthin (AST) is a dark red carotenoid with antioxidant, anti-inflammatory, neuroprotective, and antiapoptotic effects. This study investigated the ameliorative impact of AST against FPN-induced brain damage in rats. Thirty-two adult Wistar rats were allocated into four groups (n = 8): Control, AST (20 mg/kg bwt/day), fipronil (FPN) (20 mg/kg bwt/day), and AST + FPN group. Acetylcholine (ACh), dopamine, malondialdehyde (MDA), and proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and inflammatory cytokine cyclooxygenase-2 (COX2) levels were enhanced in the FPN-administered group relative to the control group. In addition, a substantial reduction of acetylcholine esterase (AchE), gamma-aminobutyric acid (GABA), serotonin, reduced glutathione (GSH) levels, catalase (CAT), and total superoxide dismutase (T-SOD) enzyme activities were determined. FPN induced histopathological alterations in the cerebral and cerebellar tissues. Likewise, the histomorphometric image analysis of H and E-stained tissue sections was constant with FPN-induced neurotoxicity. Immunohistochemically, an intense positive immunohistochemical staining of apoptotic marker caspase-3 and astrocytes activation marker glial fibrillary acidic protein (GFAP) in the examined tissues was noticed. Inversely, the simultaneous administration of AST partially attenuated FPN impacts, ameliorating the severity of FPN-induced neuronal damage. These results were also established with the molecular docking findings. It could be suggested that AST has antioxidant, anti-inflammatory, and anti-apoptotic capabilities against FPN-induced neuronal damage via suppression of oxidative stress and pro-inflammatory cytokines, preservation of the neurotransmitters, and the cerebral and cerebellar histoarchitectures.

Prospective comparison of 18F-PSMA-1007 PET/CT and MRI with histopathology as the reference standard for intraprostatic tumour detection and T-staging of high-risk prostate cancer.

To prospectively compare the ability of 18F-PSMA-1007 PET/CT and whole-body MRI (WBMRI) with DWI to detect prostate cancer (PCa) lesions and assess their local stage. Additionally, to evaluate the correlation between PSMA uptake on PET/CT and PSMA expression as assessed by immunohistochemistry. Men with newly diagnosed unfavourable intermediate or high-risk PCa underwent 18F-PSMA-1007 PET/CT and WBMRI with DWI before robot-assisted laparoscopic prostatectomy. Diagnostic accuracy for intraprostatic tumour localization, seminal vesicle invasion (SVI), and extraprostatic extension (EPE) was evaluated using whole-mount prostatectomy specimens as the reference standard. SUVmax was compared with immunohistochemical PSMA staining intensity quantified using QuPath software. 19 patients with 39 intraprostatic lesions in histopathology were included. The overall lesion detection rates for PET/CT were 84.6% and 82.1% for two independent readers, compared to 74.4% and 46.2% for MRI readers. The detection rates of index lesions were 94.7% for PET/CT and 74.0-84.0% for MRI, whereas those of non-index lesions were 70.0-75.0% for PET/CT and 20.0-65.0% for MRI. For detecting EPE, AUC values were 0.500-0.591 for PET/CT and 0.648-0.682 for MRI. For detecting SVI, AUC values ranged from 0.629 to 0.700 across both modalities. SUVmax showed a weak correlation with immunohistochemical expression of PSMA multiplied by lesion diameter (Spearman's ρ = 0.427, p = 0.013). Lesion diameters measured using 30% and 40% of SUVmax, as well as prostate SUVbackground x2, showed the closest agreement with histopathological measurements. 18F-PSMA-1007 PET/CT demonstrated high sensitivity in localizing intraprostatic carcinoma lesions but seemed inferior to WBMRI in detecting EPE. PSMA uptake appears to depend on both PSMA expression and lesion size. These findings highlight the complementary roles of PET/CT and MRI in the detection and tumor staging of PCa. Clinicaltrials.gov ID: NCT03537391. Registered 25 May 2018.

High Glycolysis and Lipid Metabolism Status Predicts Poor Prognosis in Colorectal Cancer Patients.

The prognosis of patients with stage III colorectal cancer (CRC) shows significant variations. The purpose of this study was to investigate the role of key regulatory proteins in glycolysis and lipid metabolism for the prognostic evaluation of stage III CRC patients. Utilizing the Cancer Genome Atlas (TCGA) database, we analyzed the expression of various key regulatory genes in glycolysis and lipid metabolism pathways in CRC, as well as the relationship between gene expression levels and overall survival. We selected the top two key genes exhibiting differential expression patterns in glycolysis and lipid metabolism, namely, glucose transporter type 1 (GLUT1), pyruvate kinase M2 (PKM2), fatty acid synthase (FASN), and stearoyl-CoA desaturase 1 (SCD1), as targets for subsequent exploration. We analyzed the effects of GLUT1, PKM2, FASN, and SCD1 on the proliferation, migration, and drug sensitivity of CRC cells in vitro. These proteins were detected by immunohistochemistry (IHC) in the clinical tissues of stage III CRC patients. Based on the intensity of IHC staining for GLUT1, PKM2, FASN and SCD1, the cumulative score from these 4 target proteins for each sample was calculated (score range from 0 to 8). The relationships between high (scores of 6-8) or low (scores of 0-5) expression of glycolysis and lipid metabolism molecules and the clinicopathological characteristics, and survival of patients were analyzed. The expression disparities of the GLUT1, PKM2, FASN, and SCD1 genes were the most prominent between tumor and normal tissues. Overexpression of GLUT1, PKM2, FASN, or SCD1 significantly promoted CRC cell growth and migration, as evidenced by CCK-8, colony formation, and Transwell assays. Exogenous introduction of GLUT1, PKM2, FASN, or SCD1 increased oxaliplatin IC50 values, enhanced cell survival, and reduced early apoptosis in CRC cells exposed to oxaliplatin. High glycolysis and lipid metabolism status were associated with poor tumor differentiation, vascular or nerve invasion, and shorter overall survival. The status of glycolysis and lipid metabolism was an independent prognostic factor for stage III CRC patients. High glycolysis and lipid metabolism status are correlated with a poor prognosis in patients with stage III colorectal cancer.

Evaluation of the Effect of Platelet-Rich Plasma Injection After Recurrent Nerve Injury in Rats.

Injuries to the recurrent laryngeal nerve (RLN) that may occur during thyroidectomy cause hoarseness, dysphagia, and dyspnea. Even if the injured nerve can be repaired surgically, it heals slowly and not completely. Platelet-rich plasma (PRP) is obtained by centrifuging blood taken from the human body. PRP accelerates the healing of the injured nerve due to the many growth factors it contains. The aim of this study is to investigate the therapeutic effectiveness of PRP and assess surgical repair after RLN injury. Twenty-eight male Wistar albino rats were used in this study. We divided the rats into four groups, with seven animals in each group. Group 1: RLN was cut and primary repair was performed. Group 2: RLN was cut but not repaired. Group 3: RLN was cut, primary repair was performed, and PRP was injected. Group 4: RLN was cut, and PRP was injected without repair. Laryngoscopy and electromyography (EMG) were conducted before and after the procedure. For histopathological evaluation, parameters such as Schwann cell count, axon damage, and immunohistochemical staining intensity of Ki-67 cell proliferation marker were examined. The highest amplitudes in EMG were seen in Group 3 rats at the third- and sixth-week postoperatively (p < 0.05). Regarding histopathological evaluation, Schwann cell count, and Ki-67 immunohistochemical staining were primarily observed in Group 3 rats (p < 0.05). Axonal damage and cytoplasmic vacuolization were minimally detected in Group 3 rats (p < 0.05). In our experimental rodent model, PRP injection increased the Schwann cell count and cell proliferation rate in the injured RLN area by promoting the healing of nerve axons.

Effectiveness of Biosynthesized Zinc Oxide Nanoparticles Derived From Coriandrum sativum: A Novel Nanomedicine Strategy Against Coccidiosis.

Nanomedicine has emerged as a promising solution for treating various diseases. This study inspected the invivo anticoccidial efficacy of biosynthesized zinc oxide nanoparticles (Bio-ZnNPs) from Coriandrum sativum leaves in mice infected with Eimeria papillata. Bio-ZnNPs were described using transmission electron microscopy (TEM), UV-visible spectroscopy, and x-ray diffraction (XRD). Five groups of mice were applied: a control group (uninfected), a Bio-ZnNPs group (uninfected, receiving 50 mg/kg of Bio-ZnNPs), an infection group (untreated, infected with 103 sporulated oocysts), an INF + Bio-ZnNPs group (infected, treated with Bio-ZnNPs, 50 mg/kg), and an INF + AC group (infected, treated with amprolium, 120 mg/kg). The treatments lasted for 5 days. Bio-ZnNPs were spherical, averaging 35.79 ± 7.47 nm in diameter, with a peak absorption at 300.06 nm and four diffraction peaks in the XRD spectrum. An increased excretion of oocysts was found in the INF mice group. Notable jejunal histopathological anomalies were marked by an elevation in total endogenous parasitic oocysts. There was also an increase in immunohistochemical (IHC) staining intensity of caspase-3 and enhanced antioxidant and apoptotic activity. Treatment with Bio-ZnNPs reduced oocyst excretion, improved jejunal mucosal status, and significantly decreased total parasitic stages. Post-treatment, caspase-3 levels and its IHC expression decreased, and jejunal GPX levels declined. Apoptotic gene expression showed a significant downregulation of caspase-3 and BAX, with BCL2 upregulation. Overall, Bio-ZnNPs effectively suppressed E. papillata proliferation in infected mice, suggesting their potential utility in the treatment of eimeriosis.

Identification and Clinical Validation of High HSP60 Expression Predicts Poor Prognosis in Patients with Ovarian Cancer.

This study aimed to investigate the clinical significance of heat shock protein 60 (HSP60) expression in ovarian cancer and evaluate its correlation with patient survival outcomes. A total of 260 ovarian cancer patients diagnosed between 2017 and 2019 were enrolled. Immunohistochemistry was performed to assess HSP60 expression in tumor tissues. Patients were categorized into high- or low-HSP60 expression groups based on immunohistochemical staining intensity. The correlation between HSP60 expression status and the clinicopathological features of ovarian cancer patients was analyzed. Kaplan-Meier survival curves and Cox regression models were utilized to evaluate overall survival and disease-free survival. HSP60 expression was significantly higher in ovarian cancer tissues compared to normal ovarian tissues. High HSP60 expression was associated with larger tumor size, advanced FIGO stage, and increased lymph node metastasis. Patients with high HSP60 expression exhibited significantly shorter overall survival and disease-free survival than those with low expression. Multivariate Cox analysis identified HSP60 as an independent prognostic factor for both overall survival and disease-free survival. High HSP60 expression is associated with poor prognosis and aggressive tumor characteristics in ovarian cancer. HSP60 may serve as a valuable biomarker for prognosis and a potential therapeutic target. Further randomized clinical trials are warranted to explore its role in ovarian cancer progression and treatment strategies.

PRAME Staining of Adnexal Lesions and Common Skin Cancer Types: Biomarker with Potential Diagnostic Utility.

PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen first identified in tumor-reactive T-cell clones derived from a patient with metastatic melanoma. Immunohistochemistry (IHC) for PRAME is useful for diagnostic purposes to support a suspected diagnosis of melanoma. Anecdotally, PRAME has been observed to stain sebaceous units in glands in background skin. We examined the expression of PRAME in adnexal lesions and common skin cancers to determine whether it is of potential diagnostic utility in supporting the differentiation between sebaceous and non-sebaceous lesions. IRB approval from Mount Sinai Medical Center (MSMC) was obtained. This is a single-center retrospective cohort analysis over a ten-year period (1 January 2012, and 31 December 2023). We used the pathological database of skin lesions, including sebaceous, sweat gland, and follicular lesions, in addition to basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs), from 81 patients who underwent shave/punch biopsies or surgical excisions. We evaluated the IHC staining percentage positivity and intensity for PRAME. Staining intensity was subcategorized into negative, weak, moderate, and strong, whereas expression percentage positivity was subcategorized into 0%, 1-25%, 26-50%, 51-75%, and 76-100%. Most sebaceous versus non-sebaceous lesions exhibited cytoplasmic staining of moderate to strong intensity in >75% of cells. PRAME has a sensitivity and specificity of 100.0% and 86.7%, respectively, to support distinguishing between sebaceous and non-sebaceous adnexal lesions (regardless of whether they are benign or malignant). BCCs and SCCs showed weak to moderate nuclear staining for PRAME in >75% of cells. None of the 13 lesions of hair follicle origin showed any staining. A total of 26 of the 32 lesions of sweat gland origin were negative while 6 (18.75%) showed positive staining. In conclusion, we confirm the potential utility of PRAME for supporting the distinction between sebaceous and non-sebaceous adnexal lesions on one hand, and on the other, distinguishing BCC and SCC that may show nuclear staining from sebaceous carcinoma that shows cytoplasmic staining.

Use of the immunohistochemical marker HBME-1 to optimize the diagnosis of follicular carcinomas

Background. In clinical practice, there are often patients with a diagnosis of benign follicular adenoma (FA) and follicular tumor with an uncertain malignant potential, whose diagnoses must be changed to malignant processes due to relapse or metastasis observed over time. The purpose of the study was to optimize the differential diagnosis of FA and follicular thyroid carcinoma (FTC) using the immunohistochemical (IHC) marker HBME-1 on the histological material of patients who were operated for nodular goiter with a cytological conclusion corresponding to the Bethesda categories III–V. Materials and methods. One hundred and twenty-four patients underwent surgery for follicular nodular neoplasms with a cytological conclusion according to Bethesda Gray zone and had a histological diagnosis of FTC in 23 ca­ses (18.55 %) and FA in 101 cases (81.45 %). IHC analysis was performed using mouse monoclonal antibodies against human HBME-1 (Bio SB, USA). Evaluation criteria corresponded to the strength of staining (from 0 to 3). Results. According to the intensity of IHC staining, there was an increase in the diagnosis of FTC (p &lt; 0.05): 0 — 0 %, 1 — 12.7 %, 2 — 25 %, 3 — 36.4 %. Histopathological findings were reexamined for FA that had the strongest staining reaction: in 3 (2.97 %) cases, the diagnosis was changed to FTC. Accor­dingly, the total number of detected FTC increased to 26 (20.96 %). These cases showed a good positive result regarding IHC study with HMBE-1 on FA preparations for a differential diagnosis between FA and FTC. Conclusions. HBME-1 can serve as an IHC marker for the differential diagnosis of FA and FTC in cases considered as FA, as after the study, the diagnosis was changed to FTC in 2.97 % of patients.

Chitosan-glucose derivative membrane obtained by Maillard reaction improves cartilage repair in a rabbit model

BackgroundTreatment of articular cartilage injury remains a challenging clinical problem in orthopedics. Chitosan-derived biomaterial could be a potential adjuvant treatment to improve cartilage repair. In the current study, we examined the effects of two potential chitosan-derived materials on cartilage regeneration of osteochondral defects in rabbits.MethodsAn osteochondral defect was created over the rabbit knee and treated using three approaches: group A received no material (n = 24), group B received chitosan membranes with glucose absorption (CGA; n = 25), and group C received chitosan-glucose derivative membranes obtained via the Maillard reaction (CGMR; n = 25). Cartilage repair over the osteochondral defect was analyzed 12 weeks post-surgery via histological analysis, immunostaining, and reverse transcription-qualitative polymerase chain reaction (RT-qPCR) for type-I and type-II collagen mRNA.ResultsAccording to histological analysis, CGMR-treated defects showed significantly improved modified O’Driscoll scoring when compared with no material- and CGA-treated defects (20.9 ± 4.3 vs. 13.00 ± 2.5 and 17.7 ± 4.6, p < 0.001). Moreover, group C exhibited higher intensity of type-II collagen immunohistochemical staining over the regenerated cartilage than groups A and B, along with increased expression of type-II collagen mRNA by RT-qPCR.ConclusionsCGMR might improve cartilage regeneration in osteochondral defects.

Aberrant positivity for BCOR immunohistochemistry in merkel cell carcinoma - a potential diagnostic pitfall

BackrgoundMerkel cell carcinoma (MCC) is a rare, aggressive primary cutaneous neuroendocrine carcinoma, frequently associated with clonal Merkel cell polyomavirus integration. MCC can pose significant diagnostic challenges due to its diverse clinical presentation and its broad histological differential diagnosis. Histologically, MCC presents as a small-round-blue cell neoplasm, where the differential diagnosis includes basal cell carcinoma, melanoma, hematologic malignancies, round cell sarcoma and metastatic small cell carcinoma of any site. We here report strong aberrant immunoreactivity for BCOR in MCC, a marker commonly used to identify round cell sarcomas and other neoplasms with BCOR alterations.MethodsBased on strong BCOR expression in three index cases of MCC, clinically mistaken as sarcoma, a retrospective analysis of three patient cohorts, comprising 31 MCC, 19 small cell lung carcinoma (SCLC) and 5 cases of neoplasms with molecularly confirmed BCOR alteration was conducted. Immunohistochemical staining intensity and localization for BCOR was semi-quantitatively analyzed.ResultsThree cases, clinically and radiologically mimicking a sarcoma were analyzed in our soft tissue and bone pathology service. Histologically, the cases showed sheets of a small round blue cell neoplasm. A broad panel of immunohistochemistry was used for lineage classification. Positivity for synaptophysin, CK20 and Merkel cell polyoma virus large T-antigen lead to the diagnosis of a MCC. Interestingly, all cases showed strong positive nuclear staining for BCOR, which was included for the initial work-up with the clinical differential of a round cell sarcoma. We analyzed a larger retrospective MCC cohort and found aberrant weak to strong BCOR positivity (nuclear and/or cytoplasmic) in up to 90% of the cases. As a positive control, we compared the expression to a small group of BCOR-altered neoplasms. Furthermore, we investigated a cohort of SCLC as another neuroendocrine neoplasm and found in all cases a diffuse moderate to strong BCOR positivity.ConclusionsThis study demonstrates that neuroendocrine neoplasms, such as MCC and SCLC can express strong aberrant BCOR. This might represent a diagnostic challenge or pitfall, in particular when MCC is clinically mistaken as a soft tissue or a bone sarcoma.

Dexamethasone-loaded PLGA porous microspheres used as 3D-bioprinted bioink promote tissue-engineered cartilage regeneration

The current investigation focused on fabricating and thoroughly analyzing porous microspheres (MPs) comprising poly (lactic-co-glycolic acid, PLGA) encapsulating dexamethasone, by utilizing the water-in-oil-in-water (W/O/W) emulsion solvent evaporation technique. The meticulous characterization encompassed various parameters, including particle size distribution, morphology, and polymer integrity, all of which exhibited consistent attributes across the MPs populations. The analysis of drug content and release kinetics revealed a sustained and controlled liberation of dexamethasone, characterized by an initial burst release within the initial 3 days followed by a gradual, protracted release profile. Subsequent in vivo assessments further validated the potent anti-inflammatory effects of the dexamethasone-loaded PLGA MPs, as evidenced by notable reductions in CD86 immunohistochemical staining intensity, along with the change in membrane thickness. Additionally, the MPs demonstrated an impressive capability to support the maintenance of cartilage phenotype, as indicated by the augmented secretion of cartilage matrix components and elevated glycosaminoglycan content. &amp;nbsp;

Mesothelin promotes the migration of endometrioid carcinoma and is associated with the MELF pattern

Mesothelin promotes the migration of endometrioid carcinoma and is associated with the MELF pattern

Deciphering the prognostic value of FGFR2b/2c isoform expression levels in advanced esophagogastric cancer through whole-transcriptome sequencing.

53 Background: Fibroblast growth factor receptor (FGFR) isoform switching from FGFR2b to FGFR2c has been implicated in epithelial-to-mesenchymal transition and enhanced invasive potential in various malignancies and potentially be involved in resistance mechanisms to FGFR2b targeted therapy (e.g., bemarituzumab). This study sought to investigate the potential of respective FGFR2 isoform expression evaluation utilizing whole-transcriptome sequencing (WTS) and its correlation with therapeutic efficacy and clinical outcomes in patients with advanced G/GEJ cancer. Methods: Patients with advanced G/GEJ cancer who received systemic therapy at the National Cancer Center Hospital East between May 2021 and September 2023, and were enrolled in the immunological profiling study (UMIN000019129) and MONSTAR-SCREEN-2 (UMIN000043899) were included. FGFR2 protein and RNA expression were assessed via immunohistochemistry (IHC) and WTS with Caris MI Profile, respectively. FGFR2b and FGFR2c isoforms were distinguished by the detection of their respective splice junctions using WTS data. The FGFR2b/(2b+2c) ratio was stratified into FGFR2b-dominat and -containing subtypes using the median value. The log-rank test was used to discern differences in overall survival (OS). Results: Among the 129 patients, 21 (16.3%) exhibited FGFR2-positivity by IHC (staining intensity 2+ or 3+≥1% membranous staining of tumor cells), and a significant correlation was observed between FGFR2 IHC status and RNA expression levels (TPM: transcript per million) in WTS (FGFR2-positive, median TPM=11.1; FGFR2-negative, median TPM=4.7; P=0.002). The distribution of FGFR2b/(2b+2c) ratio (median, 0.89) was: 10.9% (0≤x&lt;0.2), 5.4% (0.2≤x&lt;0.4), 7.0% (0.4≤x&lt;0.6), 12.4% (0.6≤x&lt;0.8), 43.4% (0.8≤x&lt;1.0), and 20.9% (x=1.0). Among 58 patients treated with 1st line therapy, no significant difference in treatment response was observed between the FGFR2b-dominat (67.9%) and FGFR2c-containing subtypes (53.3%) (P=0.259). Overall, the FGFR2c-containing subtype was associated with significantly shorter OS compared to the FGFR2b-dominat subtype (median OS, 10.1 vs. 17.9 months; hazard ratio, 1.72; 95% confidence interval, 1.03-2.86; log-rank P=0.037). Conclusions: Our findings underscore the substantial concordance between IHC and WTS analysis of FGFR2, the feasibility of distinguishing FGFR2 isoforms using WTS, and the prognostic implications of FGFR2c isoform in patients with advanced G/GEJ cancer. Further analysis is warranted in patients receiving FGFR2b targeted treatment.

Semiquantitative assessment of phosphatase and tensin homolog value with immunohistochemistry in colorectal cancer.

Colorectal cancer has emerged as a concerning health problem, ranking the third most common form of cancer in both men and women. The phosphatase and tensin homologue (PTEN) protein is widely known for its role as an inhibitor of the phosphatidylinositol 3-kinase/protein kinase-B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway, playing a major role inhibiting tumor development. Previous studies investigated the role of this protein in the PI3K pathway and how it affected colorectal cancer. However, a standardized cut-off value for PTEN expression has not been established. Immunohistochemistry was used in examining PTEN. The staining grade ranging from 0 to 3 was then multiplied by the number of 100 cancer cells counted, with total score between 0 and 300. In this study, receiver operating characteristic (ROC) curve was employed to determine the expression cut-off value for PTEN in colorectal cancer. This study showed statistically significant results (P < 0.001) in either tumor or non-tumor tissues by using the ROC curve with a cut-off value of 199.0. This study also revealed significant correlation between nodal status with PTEN (P = 0.008) and stage with PTEN (P = 0.019) with sensitivity 0.753 and specificity 0.728. Semiquantitative assessment with cell counting multiplied by color intensity is a good method in determining PTEN expression. The use of immunohistochemical staining intensity and cell scoring with ROC cut-off is effective to elaborate the effects of PTEN in colorectal cancer (PTEN value > 199.0 was classified as strong and ≤ 199.0 as weak).

Immunoreactivity of LMO7 and other molecular markers as potential prognostic factors in oropharyngeal squamous cell carcinoma

BackgroundDespite the better prognosis associated with human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC), some patients experience relapse and succumb to the disease; thus, there is a need for biomarkers identifying these patients for intensified treatment. Leucine-rich repeats and immunoglobulin-like domain (LRIG) protein 1 is a negative regulator of receptor tyrosine kinase signaling and a positive prognostic factor in OPSCC. Studies indicate that LRIG1 interacts with the LIM domain 7 protein (LMO7), a stabilizer of adherence junctions. Its role in OPSCC has not been studied before.MethodsA total of 145 patients diagnosed with OPSCC were enrolled. Immunohistochemical LMO7 expression and staining intensity were evaluated in the tumors and correlated with known clinical and pathological prognostic factors, such as HPV status and LRIG1, CD44, Ki67, and p53 expression.ResultsOur results show that high LMO7 expression is associated with significantly longer overall survival (OS) (p = 0.044). LMO7 was a positive prognostic factor for OS in univariate analysis (HR 0.515, 95% CI: 0.267–0.994, p = 0.048) but not in multivariate analysis. The LMO7 expression correlated with LRIG1 expression (p = 0.048), consistent with previous findings. Interestingly, strong LRIG1 staining intensity was an independent negative prognostic factor in the HPV-driven group of tumors (HR 2.847, 95% Cl: 1.036–7.825, p = 0.043).ConclusionsWe show for the first time that high LMO7 expression is a positive prognostic factor in OPSCC, and we propose that LMO7 should be further explored as a biomarker. In contrast to previous reports, LRIG1 expression was shown to be an independent negative prognostic factor in HPV-driven OPSCC.

Identification of HER2 ultra-low based on an artificial intelligence (AI)-powered HER2 subcellular quantification from HER2 immunohistochemistry images.

1115 Background: HER2-targeted antibody-drug conjugates (ADCs) can effectively target tumor cells even in HER2-low breast cancers (1+ and 2+/no amp). We developed an AI-based whole-slide image (WSI) analyzer for immunohistochemistry (IHC)-stained slides, exploring a cutoff differentiating between true HER2 negative versus HER2 ultra-low. Methods: An AI-based IHC WSI analyzer was developed using 6,188 WSIs stained by 19 types of IHC including HER2 and from various cancers, annotated by 153 pathologists. The model predicts cells as tumor cells or non-tumor cells and generates HER2 expression continuous score (HER2ecs, 0-100%), quantifying IHC staining intensity of membrane, nucleus, cytoplasm, and membrane+cytoplasm separately for each cell. A total of 401 HER2 IHC WSIs were acquired from Samsung Medical Center (n=275), Kyung Hee University Hospital (n=78), and Commercial Biobank (n=48), scored as 0 (n=347) and 1+ (n=54) by pathologists. Results: In total, 67,169,114 tumor cells and 119,113,886 non-tumor cells from 401 WSIs were successfully evaluated. Comparing HER2 score 0 versus 1+ by pathologists, mean HER2ecs of nucleus (1.7% vs 4.1%), cytoplasm (6.5% vs 12.9%), and membrane (3.6% vs 8.2%) were comparable. In HER2 score 0 WSIs, membrane HER2ecs of tumor cell was 3.6 ± 7.8% (mean ± standard deviation), while that of non-tumor cell was 0.9 ± 2.9% (p &lt;0.001 vs. tumor cell), suggesting, even in HER2 score 0 cases, tumor cell exhibits HER2 expression beyond non-specific HER2 staining intensity. Based on our internal validation dataset, membrane-specific HER2ecs of 6.0% was the optimal threshold discriminating HER2 no-staining cells versus HER2 incomplete and faint/barely perceptible cells (AI-HER2-weak) per ASCO/CAP guideline. Among HER2 score 0 by pathologists, when applying this cutoff, the proportion of WSIs showing AI-HER2-weak or higher membrane-specific HER2ecs in over 10% of all tumor cells was 23.6% (82/347). Interestingly, that of WSIs among HER2 score 1+ by pathologists was 51.9% (28/54), which is comparable to 52.3%, objective response rate of trastuzumab deruxtecan in HER2-low patients in DESTINY-Breast04. Conclusions: AI-powered subcellular-level analysis enables identification of 23.6% HER2 ultra-low WSIs among HER2 score 0 by pathologist, which is defined by AI-HER2-weak or higher membrane-specific HER2 expression ≥ 10%. Clinical validation of the AI-powered HER2 ultra-low definition is warranted.

Assessment of Mammalian Target of Rapamycin Pathway Activation in Basal Cell Carcinoma as a New Therapeutic Approach.

Targeting the mammalian target of rapamycin (mTOR) pathway represents a potentially novel approach to treat basal cell carcinoma (BCC), but activation of this pathway has not been well described in human BCCs. The purpose of this study was to assess whether mTOR pathway activation occurs in BCCs (both sporadic and syndromic) and report a case of a patient with Gorlin syndrome (GS) whose clinically suspicious BCCs responded to mTOR inhibition through topical sirolimus treatment. After Stanford Institutional Review Board Approval, archived BCCs from patients with GS (n = 25), sporadic BCCs (n = 35), and control tissues were subjected to immunohistochemical analysis for the activation of mTOR pathway, and immunohistochemical staining intensity was evaluated by a dermatopathologist. BCCs (compared with normal skin) had elevated levels of eIF4EBP1 ( Padjusted = 0.0336), which is downstream of mTOR. a serine/threonine kinase Phospho-(AKT), which interacts with mTOR, was also significantly elevated (perinuclear: Padjusted < 0.0001; cytoplasmic: Padjusted = 0.0021). When off-label topical 1% sirolimus was used on a pediatric patient with GS, we noted reduction of new BCC development and decreased size of existing neoplasms clinically suspicious for BCCs. This treatment was well tolerated after 2 years of continuous use, with no other treatments needed during this period. Topical sirolimus is a promising therapeutic candidate against both sporadic and GS-associated BCC. Multicenter, prospective studies are needed to understand the efficacy and safety of topical mTOR inhibitors in BCC treatment, and ascertain whether the immunohistochemical markers downstream of mTOR could have predictive value in identifying BCCs most likely to respond to topical mTOR inhibitors, such as sirolimus.