High Proliferation Index Research Articles

FGF/FGFR1 system in paired breast tumor-adjacent and tumor tissues, associations with mammographic breast density and tumor characteristics.

Mammographic breast density (MBD) is an established breast cancer risk factor, yet the underlying molecular mechanisms remain to be deciphered. Fibroblast growth factor receptor 1 (FGFR1) amplification is associated with breast cancer development and aberrant FGF signaling found in the biological processes related to both high mammographic density and breast cancer microenvironment. The aim of this study was to investigate the FGF/FGFR1 expression in-between paired tumor-adjacent and tumor tissues from the same patient, and its associations with MBD and tumor characteristics. FGFR1 expression in paired tissues from 426 breast cancer patients participating in the Karolinska Mammography Project for Risk Prediction of Breast Cancer (KARMA) cohort study was analyzed by immunohistochemistry. FGF ligand expression was obtained from RNA-sequencing data for 327 of the included patients. FGFR1 levels were differently expressed in tumor-adjacent and tumor tissues, with increased FGFR1 levels detected in 58% of the tumors. High FGFR1 expression in tumor tissues was associated with less favorable tumor characteristics; high histological grade (OR=1.86, 95% CI 1.00-3.44), high Ki67 proliferative index (OR=2.18, 95% CI 1.18-4.02) as well as tumors of Luminal B-like subtype (OR=2.56, 95%CI 1.29-5.06). While no clear association between FGFR1 expression and MBD was found, FGF ligand (FGF1, FGF11, FGF18) expression was positively correlated with MBD. Taken together, these findings support a role of the FGF/FGFR1 system in early breast cancer which warrants further investigation in the MBD-breast cancer context.

Expression of BRCA1 by immunohistochemistry and its association with ER, PR, Her2neu status in infiltrative ductal carcinoma of breast.

Breast cancer is a heterogeneous disease, which differs in its clinical behaviors and responses to treatment and outcome. The prognosis of breast cancer depends on histopathological parameters and molecular subtypes. Among more than 300 genes, which are involved in the pathogenesis of breast cancer tumor suppressor gene such as BRCA1 is known to play a significant role in hereditary cancers. However, its role in sporadic cases of infiltrating ductal carcinoma is yet to be established. To evaluate the expression of BRCA1 in infiltrative ductal carcinoma and to analyze the association of BRCA1 with histopathological parameters and estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor2 (Her2) neu expression. This was a laboratory-based exploratory study in which 56 patients with infiltrative ductal carcinoma who underwent radical mastectomy from October 2019 to July 2021 were included. Patients with chemotherapy and radiotherapy, trucut biopsies, and incomplete patient details were excluded. Immunostaining for BRCA1 was performed. Individual clinicopathological parameters were compared with the BRCA1 mutation. Statistical analysis was done using SPSS 22. A P value of < 0.05 was considered statistically significant. Among 56 cases of IDC, 18 cases (32.1%) showed BRCA1 mutation. BRCA1 mutation was associated with postmenopausal age, larger tumor size, lower tumor grade, and higher tumor staging. When we analyzed the biomarkers with BRCA1 mutation, it showed a negative association with ER, PR, and Her2 neu and a high Ki67 proliferation index. No family history of breast carcinoma was seen in 34/56 patients where history was available. Our study showed BRCA1 mutation in 32.1% and associated with postmenopausal age group, larger tumor size, and higher staging and negative hormonal status of breast carcinoma.

Expression of EZH2 and H3K27me3 predicts tumor biology of urothelial carcinoma.

Enhancer of zeste homolog 2 (EZH2) is one of the major epigenetic modifiers involved in the transcriptional repression of target genes through trimethylation of H3K27 (lysine 27 residue of histone H3). Deregulated expression of both EZH2 and H3K27me3 has been implicated in the biological behavior and prognostic outcome of various malignancies. To assess the role of EZH2 and H3K27me3 in the carcinogenesis of urothelial carcinoma of urinary bladder. One hundred fifty consecutive urothelial carcinoma cases of urinary bladder (54.7% high-grade) were included in this study. Immunohistochemical analysis for EZH2 and H3K27me3 was performed on whole tissue sections. A multiplication score obtained by multiplying staining intensity and proportion of positively stained neoplastic cells was used for assessment. EZH2 showed a significant correlation with the tumor grade and lamina propria invasion (p < 0.001). The cases with high EZH2 expression showed a significantly high proliferative index (Mean- 32.7%; p < 0.001). In contrast, negative and low expression of H3K27me3 was significantly more common in high-grade cases (p = 0.006). The expression of H3K27me3 was significantly associated with lamina propria (p = 0.01) and deep muscle invasion (p = 0.007). EZH2 showed a significantly higher expression in the high-grade invasive areas as compared to the high-grade non-invasive areas of the same tumor (p = 0.03). This study establishes an important role of the key epigenetic regulators EZH2 and H3K27me3 in the pathobiology of urothelial carcinomas. Strong expression of EZH2 and weak expression of H3K27me3 are associated with higher grade, proliferative index and invasive behavior.

Primary cutaneous blastic marginal zone lymphoma: A comprehensive clinical, light microscopic, phenotypic and cytogenetic appraisal

BackgroundPrimary cutaneous marginal zone lymphoma (PCMZL) is a form of indolent lymphoproliferative disease where the disease is largely a cutaneous confined process. It is typically a neoplasm composed of post germinal small B-cells and light chain restricted plasma cells in a background of reactive T-cell hyperplasia and benign germinal centers. Rarely a significant degree of large cell infiltration occurs warranting the categorization as blastic marginal zone lymphoma. Materials and methodsWe reviewed our data base over a time period of 2016 to 2022 for cases diagnosed as blastic MZL. Twelve cases were identified. The clinical records and pathological data were reviewed. ResultsNine of the cases represented de novo forms of blastic MZL while in three cases there was a prior history of MZL. Multifocal cutaneous disease was not uncommon and one quarter of the cases had evidence of extracutaneous dissemination. All patients except three achieved remission with varied therapeutic interventions depending on the extent of the disease ranging from conservative re-excision to chemotherapy. No patient died from lymphoma. Light microscopically, there was evidence of a background of conventional MZL in the majority of cases. The large cell component was typically characterized by multiple micronodular aggregates throughout the dermis although in three cases there was a striking diffuse large cell component as the dominant infiltrate. Phenotypically, a third of the cases showed either CD5 or CD23 positivity amidst neoplastic B cells. Significant staining for BCL-2 was noted in the majority of cases tested while extensive MUM-1 positivity was observed in half of the cases tested. Kappa or lambda light chain restriction was seen in most. The Ki67 proliferation index exceeded 30 % in all cases. There was C-MYC positivity in two cases. While most cases did not detect cytogenetic abnormalities, one case had multiple cytogenetic hits that are associated with diffuse large B cell lymphoma. Next generation sequencing showed a Ten-eleven translocation 2 mutation in the earlier biopsy prior to transformation and in the later biopsy after transformation along with an additional B2M mutation in the transformed biopsy. Both types of mutations are very uncommon but held to contribute to tumor progression in the setting of diffuse large B cell lymphoma. ConclusionBlastic MZL is associated with a more aggressive clinical course. Even when there is disseminated disease patients while not always cured did not have a fatal course in this series. The light microscopic findings are reproducible. The background of MZL, identification of larger cells in significant numbers without a follicle center phenotype, at times expressing CD5 or CD23 with variable positivity for MUM1, BCL-2 and C-MYC and a high proliferation index define the pathology in most. Certain cytogenetic abnormalities and genetic mutations implicated in large cell transformation into a diffuse large B cell lymphoma are seen in blastic MZL with earlier biopsies prior to transformation potentially harboring at risk genetic mutations.

Expression of Ki-67 antigen in various histological types of BASAL CELL CARCINOMA

Background : Basal cell carcinoma (BCC) is the most commonest malignant tumor of the skin especially fair-skinned individuals makes up roughly 70–80% of all malignant tumors. In certainty, this malignancy had a variety of many tumor sub-types, through a distinct histo-morphological appearance and biological function. Unlike other tumors, usually has an affirmative clinical course, developing slowly and just locally. Antibodies against “Ki-67 antigen” or proliferating Cell nuclear antigen (PCNA) are typically used in immunohistochemistry to detect the proliferative activity (growth fraction) of malignancies. Aim of the study : This study aims to examine the tumor cells' proliferative status utilizing an immunohistochemistry analysis of the “Ki-67” antigen in skin biopsy specimens of different sub-type of BCCs. Material and methods : A Case series study enrolling 35 biopsies sampled from retrospectively selected cases of cutaneous BCCs were registered in the research. Results : Thirty five biopsy (31 Primary, 4 Recurrent)of different histological types from 35 patients (11 male, 24 female) were included. The patient’s age ranged from 36 to 75 years (with mean of 59.2 year) .In this study results showed that commonest site of involvement was the face in 71.4% of cases , and there was significant association between tumor size and the Ki67 level and grade, the study reported a non-significant association between the tumor histological subtype and Ki67 level. Discussion : The data showed that there were a wide range of ki67 level (2 to 70%) among various histological growth subtype of basal cell carcinoma, higher proliferative index indicate more malignant potential and lower proliferative index mean less aggressive tumor. So the wide range of proliferation which had been reported in the BCC illuminate the wide spectrum of biological activity of tumor.

Cell-In-Cell Structures in Early Breast Cancer Are Prognostically Valuable

Cell-in-cell (CIC) structures in breast cancer have so far been studied in a small inhomogeneous patient population, suggesting the prognostic importance of CIC. In the present study, we focused on CIC in early hormone-sensitive breast cancer. With in vitro co-culture experiments, we compared the homotypic phagocytic capacity of two breast cancer cell lines to that of primary human fibroblasts. Afterward, we studied 601 tissue specimens from 147 patients participating in an institutional accelerated partial breast irradiation (APBI) phase II trial. Both breast cancer cell lines performed non-professional phagocytosis at a higher rate than primary human fibroblasts. In this study cohort, 93.2% of the patients had T1 tumours, and 6.8% had T2 tumours. CIC was found in 61.2% of the patients, with a CIC rate ranging from <1/mm2 to 556.5/mm2 with a mean of 30.9/mm2 ± 68.4/mm2. CIC structures were prognostically favourable for local recurrence-free survival and disease-free survival. Regarding metastasis-free survival, CIC-positive patients had an unfavourable prognosis. Subgroup analysis indicated a correlation between a high proliferation index and high CIC rates. CIC had the highest prognostic value in young breast cancer patients (p = 0.004). With this study, we provide further evidence of CIC as a prognostic marker in breast cancer.

Does the number of cell forming tumor budding alter the prognostic value in invasive ductal carcinoma of breast?

The most commonly used definition for tumor budding (TB) is a single or a cell cluster of tumor cells up to 4 cells. However, there are different opinions regarding the number of cell (NOC) forming TB. It has been proven that TB is associated with poor prognostic factors in most tumors. The current study, it was aimed to investigate the prognostic value of NOC forming TB in invasive ductal carcinoma of the breast. 326 cases with the diagnosis of invasive ductal carcinoma were examined. The NOC forming TB was counted from hematoxylin and eosin stained slide under X200 magnification for each case, and scoring five different TB as 1, ≤2, ≤3, ≤4, ≤5, respectively. Receiver operating characteristic (ROC) analysis based on survival was performed for each TB value separately, and the cut-off was determined. All TB values were associated with poor outcome (p<0.001), presence of distant metastasis (p<0.001), high Ki67 proliferation index (p<0.05), advanced stage (p<0.05), presence of lymphovascular invasion (p<0.001), and metastatic axillary lymph node (p<0.001). According to ROC analysis performed to compare the predictiveness of survival, the area under the curve was similar for all TB values. TB was associated with poor prognostic parameters, and the prognostic value of TB was not affected by NOC forming TB. The NOC up to 4 cells which have been accepted for colon carcinomas, could also provide practicality in breast carcinomas.

Recurrent posterior fossa group A (PFA) ependymoma in a young child with constitutional mismatch repair deficiency (CMMRD).

Recurrent posterior fossa group A (PFA) ependymoma in a young child with constitutional mismatch repair deficiency (CMMRD).

Differential Phosphoproteomics Deciphers Physiopathology of High-Risk Mantle Cell Lymphoma

Background: Mantle cell lymphoma (MCL) is a rare adult-onset non-Hodgkin lymphoma of which genetic hallmark is CCND1 translocation with IG genes, although CCND2, CCND3 or CCNE translocations are alternatives. The median survival of patients with MCL has lately increased due to improved therapies. However, the identification of high-risk MCLs characterized by pleomorphic or blastoid appearance, high proliferation index and TP53 deletions/mutations, remains mandatory due to their highly aggressive features and dismal clinical course even with intensified regimens. Therefore, specific therapeutic approaches are still warranted for such patients, but clinical and biological data are scarce since high-risk MCLs represent only 10-20% of all MCLs. A valuable source of rare samples for clinical/translational research is archived collections of formalin-fixed paraffin-embedded (FFPE) tissue blocks which enable retrospective studies to be performed. However, FFPE samples remain challenging for omics studies, especially phosphoproteomics. Here we implemented an original process to overcome this technical roadblock and took full advantage of FFPE samples to investigate biological processes differentially involved in high-risk and classical MCL (cMCL) to highlight rationale for innovative targeted therapies for these aggressive forms. Material and methods: We employed an unbiased differential label-free phosphoproteomic approach using archived FFPE MCL samples. Briefly, archived FFPE lymph nodes conclusive for cMCL or blastoid MCL (bMCL) diagnoses were retrieved from the Biological Resource Center of Strasbourg University Hospital from 1996 to 2016. H&E slides were reevaluated, and areas enriched in tumoral cells were selected to be punched from FFPE blocks. Our original sample preparation procedure entailed 1/ xylene-based deparaffinization, 2/ protein extraction with high content SDS-based lysis buffer, 3/ sample reduction/alkylation followed by cleanup and protein digestion on carboxylate paramagnetic beads and Lys-C/trypsin proteolytic enzyme mix, 4/ phosphopeptide enrichment on the AssayMap Bravo platform (Agilent) using IMAC Fe(III)-NTA cartridges. All samples were then analyzed on a NanoElute coupled to a TimsTOF Pro mass spectrometer (Bruker). Label-free quantification was performed using MaxQuant with the human database downloaded from SwissProt in April 2022. Further bioinformatics analyses were conducted in Perseus and Prostar. Data were functionally annotated with Reactome Pathway Database and STRING for association networks. Results: For this monocentric study we retrieved 30 FFPE blocks of lymph node biopsies conclusive for MCL diagnosis, including 15 bMCL and 15 cMCL. Although FFPE samples represented a challenge to work with for phosphoproteomics, we extracted enough protein from three 2mm-punches to proceed to the phosphopeptide enrichment according to our original sample preparation procedure. Overall, we successfully identified and quantified 7,220 class I phosphosites, i.e., sites of phosphorylation for which the probability of the phosphate group localization is greater than 0.75. No significant difference was found considering neither the archived period length of FFPE blocks nor the MCL subtypes (bMCL or cMCL). Differential analysis of bMCL and cMCL identified 180 differentially phosphorylated phosphosites (p-value < 0.05) corresponding to 103 proteins. Among these phosphosites, 139 were more phosphorylated in bMCL. Functional annotation of their corresponding proteins highlighted an over-representation of proteins involved in cell cycle, DNA repair and RNA metabolism within bMCL differential phosphoproteome (Fig.1). In addition, the kinase enrichment analysis of these differentially phosphorylated proteins predicted an increased activity of cyclin-dependent kinases in bMCL compared to cMCL. Conclusion: Here we performed for the first time phosphoproteomics from archived FFPE MCL samples. Differential analysis revealed that high-risk MCLs are characterized by an increased phosphorylation of proteins involved in cell cycle and an increased activity of cyclin-dependent kinases. Further studies are mandatory to fully understand to which extent cyclin-dependent kinases could be suitable targeted therapeutic candidates for patients with high-risk MCL. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

The Age, Comorbidities and Albumin (ACA) Index Is Independently Associated with Overall Survival in Patients with Mantle Cell Lymphoma (MCL)

Background: Mantle cell lymphoma (MCL) is a biologically and clinically heterogenous disease with variable outcomes. Numerous prognostic factors have been identified, including blastoid histology, the MCL International Prognostic Index (MIPI) score, high proliferation index, TP53 deletions and/or mutations, complex karyotype, minimal residual disease positivity, and several others. Most prognostic scores do not take patient characteristics into account and require additional testing which may not be routinely available. In particular, older adults have heterogenous outcomes due to the presence of comorbidities, impaired functional and/or nutritional status which is not factored into the MIPI score. The age, comorbidities and albumin (ACA) index is a simple score that can be gleaned from the electronic health record either in real-time or retrospectively and has been validated in patients with diffuse large B cell lymphoma. We evaluated the impact of the ACA index on outcomes in patients with MCL. Methods: The Roswell Park MCL database includes all patients with MCL evaluated at Roswell Park Comprehensive Cancer Center from 2007 to 2019. A serum albumin (SA) level of <3.7 g/dL was defined as "low". The age, comorbidities and albumin (ACA) index included a low SA level, age ≥ 75 years, and a Charlson comorbidity index (CCI) score of ≥3 (representing moderate to severe comorbidity burden), with each variable when present contributing to a score of 1. A score of 0 was defined as "excellent", 1 as "good", 2 as "moderate" and 3 as "poor". The log-rank test was used to compare overall survival (OS), and the Cox-proportional hazard model was used for univariate (UVA) and multivariate (MVA) survival analyses. Results: A total of 245 patients with a median age of 64 years [interquartile range (IQR), 56-73 years] were included in the study, with a median follow-up of 90.2 months (95% CI, 75.1 - 107 months). Overall, 111 (45%) patients were age ≥65 years, 81 (33.1%) were age ≥70 years, and 44 (18.0%) were age ≥75 years; 73% of patients were male, and 11 patients had an ECOG PS ≥2. Older adults (age ≥75 years) had a greater burden of comorbidities (CCI≥3, p<0.001), and poor PS (ECOG ≥2, p=0.006) compared with younger patients. The median SA in the entire cohort was 4.30 g/dL (IQR 3.90, 4.50), with lower median levels in patients age ≥75 versus age <75 years (4.00 vs 4.35, p<0.001); however, there was no difference in frequency of patients with low SA levels (<3.7 g/dL) (19% vs 14%, p=0.4). A low SA level, CCI≥3, high or intermediate MIPIb risk, ki-67>30%, and increased age (≥75 years) were significantly associated with inferior OS. Patients who received an autologous or allogeneic stem cell transplant (SCT) in any line of therapy had improved OS (HR 0.5517, p=0.001). The ACA score could be calculated in 69.8% of patients- 58% of patients had an excellent ACA score (0), 30% of patients had a good score (1) and 9.9% of patients had a moderate score (2). Only 2 (1.2%) patients had a poor score (3). Patients with higher ACA scores had significantly inferior OS (HR 2.117, p<0.0001). In a MVA that adjusted for MIPIb risk and treatment with SCT in any line, this risk remained significant (HR 1.67, p=.007). Conclusion: A simple, patient-factor based scoring system incorporating hypoalbuminemia and the presence of comorbidities is strongly predictive of survival in patients with MCL, irrespective of disease stage, MIPIb risk score, and known molecular risk factors. Further steps include validation of the ACA index in an independent MCL cohort and incorporation of elements from the ACA index into known prognostic models to develop a new score that takes both disease biology and patient physiology into account. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract PR012: Perioperative liver biopsy captures features of the liver pre-metastatic niche and predicts metastatic outcome after pancreatic cancer resection

Abstract Pancreatic cancer (PC) has a high propensity for liver metastasis (LM), a rapidly lethal event, often seen early after resection of the primary tumor. Mounting pre-clinical evidence implicates the establishment of microenvironmental alterations in the liver or other target organs prior to emergence of clinically evident metastases, termed “pre-metastatic niches”, which is a crucial first step in metastatic progression. To determine the translational relevance of these findings, we evaluated liver biopsies from PC patients obtained during pancreatectomy to characterize the cellular, molecular and metabolic features that define the pre-metastatic niche and that may, in turn, be used as biomarkers to stratify the metastatic risk, and as therapeutic targets to interrupt the metastatic cascade. Thus, liver biopsies from 49 patients with localized PC were analyzed by transcriptomics, metabolomics, histopathology, flow cytometry, and in selected patients by single cell RNA sequencing. Patients underwent routine surveillance in a prospective manner to determine which patients developed early (&amp;lt;6 months) or late (&amp;gt;6 months) LM, extrahepatic metastasis, or remained disease-free. Eighteen patients with non-cancerous pancreatic lesions undergoing pancreatectomy were used as controls (non-PC). Single cell transcriptomics analyses and immuno-profiling revealed that, in contrast to non-PC livers, tumor-free, pre-metastatic livers from PC patients exhibited enhanced inflammation evidenced by enrichment of neutrophils, CD11B+ cells, and CD3+ T cells, as well as expansion and activation of a CD11B+ subset of NK cells. Furthermore, immune and metabolic profiling of PC pre-metastatic livers identified features that predicted future patterns of metastasis: high Ki67 proliferation index, neutrophil extracellular trap (NET) formation, and upregulated sortilin-1 gene expression, all of which correlated with earlier LM. Conversely, widespread lobular inflammation, with abundant lobular CD3+ T cells, as well as intact creatine metabolism and steatosis associated with no recurrence, isolated extrahepatic recurrence or later LM. We combined the above variables to develop a decision tree-based prediction model, which performed best for identification of early LM (AUC 0.85). Taken together, these data demonstrate that in patients with apparently localized PC, pre-treatment liver biopsies may confirm the presence of human hepatic pre-metastatic niche and reveal features that correlate with timing and patterns of recurrence (early or late LM, extrahepatic, or no recurrence), after resection of PC. These liver profiles represent novel biomarkers that may be used to provide prognostic information and guide subsequent treatment of PC. Citation Format: Linda Bojmar, Constantinos P. Zambirinis, Jonathan Hernandez, Jayasree Chakraborty, Samer Hanna, Diane Simeone, David Kelsen, Haiying Zhang, Irina R Matei, Per Sandström, Robert Schwartz, William R. Jarnagin, David Lyden. Perioperative liver biopsy captures features of the liver pre-metastatic niche and predicts metastatic outcome after pancreatic cancer resection [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer; 2022 Sep 13-16; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2022;82(22 Suppl):Abstract nr PR012.

Exploring New Prognostic Biomarkers in Mantle Cell Lymphoma: The Best RNA Based Risk Score

INTRODUCTION Mantle Cell Lymphoma (MCL) is characterized by heterogenous disease courses ranging from indolent cases, that do not require therapy for years, to highly aggressive disease with a poor prognosis despite intensive treatment regimens. This emphasizes the need for accurate prognostic biomarkers to guide risk-adapted treatment strategies. The established risk factors used in the clinic today including high Mantle Cell Lymphoma International Prognostic Index (MIPI), high Ki67 proliferation index, high MIPI-c, TP53 mutations, and blastoid morphology, only partly identify patients in need of alternative treatment. Therefore, the search for clinically fit and biologically relevant biomarkers has been intensified. This has led to the proposal of two new NanoString-based, proliferation-associated prognostic biomarkers: the mRNA-based MCL35 score (Scott et al. 2017) and the circular RNA based circSCORE (Dahl et al. 2021). Both biomarkers have shown promising prognostic potential, but a comparison has never been done before. In this study we aimed at comparing MCL35 and circSCORE in 149 patients from the uniformly treated MCL2 and MCL3 cohorts. MATERIALS AND METHODS We included 149 patients from the Nordic patient cohorts MCL2 (70 patients) and MCL3 (79 patients) with sample data on both circSCORE and MCL35 along with long-term follow up data including progression free survival (PFS) and overall survival (OS). Gene expression analysis was obtained with the NanoString nCounter technology after RNA-isolation, using samples originating from both diagnostic lymph-nodes and non-nodal tissue. We also performed individual analyses on the MCL3 cohort only to eliminate the potential risk of overfitting, since the circSCORE is trained upon the MCL2 cohort. RESULTS We report that both circSCORE (PFS log-RANK p-value 0.0048 and OS log-RANK p-value 0.0426) and MCL35 (PFS log-RANK p-value 0.0422 and OS log-RANK p-value 0.0444) in the MCL3 only survival analyses displayed equal significant, prognostic potential in stratifying high and low risk groups. These analyses also revealed inability to significantly stratify the MCL35 standard risk group from the high- and low MCL35 risk groups (figure 1). Neither continuous circSCORE nor continuous MCL35 held prognostic significance when adjusting for known high-risk factors in multivariable cox regression, where only MIPI and TP53 mutation maintained significant results in all settings, except for PFS in MCL3 only (data not shown). The categorial data were analyzed with log-likelihood (LL) using the MIPI and TP53wt (wild type) cox models as baseline models (table 1). The results showed that both circSCORE and Ki67 added significant improvement to the MIPI baseline model in the pooled, MCL2+MCL3 setting, but most interestingly, only circSCORE added significant improvement in MCL3 only for PFS (LL -97.655 vs -100.197, p-value 0.011). Neither circSCORE, MCL35 nor Ki67 added significant improvement to the TP53wt baseline model (table 1). CONCLUSION Both circSCORE and MCL35 demonstrated equal prognostic potential in separating high-and low risk groups in this study. Neither circSCORE, MCL35 nor Ki67 added significant improved value to the TP53wt baseline cox model, but circSCORE added significant improved value to the MIPI baseline cox model, even outperforming Ki67. This implies a new combined prognostic biomarker "MIPI-circSCORE", which could reflect the complexity of MCL through both clinical and biological, proliferation-associated features. However, the overall performance of the MCL35 risk score might be affected due to the inability of separating the standard risk group from the other MCL35 risk groups, the limited number of patients, and the tissue specific criteria of the MCL35 score not being met in this study. Therefore, the findings of this study should be further explored in larger MCL-cohorts to ensure stronger statistical power. Also, testing the biomarkers in MCL-relapse would be of great value to examine the prognostic potential in other settings of the heterogenic course of MCL. This is warranted to clarify the clinical future relevance of these biomarkers in supporting risk-adapted treatment regimens in patients with MCL. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

PATH-42. DIAGNOSIS AND PROGNOSTICATION OF AN ATYPICAL ADULT PRESENTATION OF DIFFUSE LEPTOMENINGEAL GLIONEURONAL TUMOR (DLGNT) BY METHYLATION PROFILING

Abstract BACKGROUND Diffuse leptomeningeal glioneuronal tumors (DLGNT) are rare central nervous system tumors first described in the 2016 WHO classification. A paucity of data exists on this tumor. This case describes an adult presentation of DLGNT, a classically pediatric tumor, and highlights the importance of genetic and epigenetic analyses in neuro-oncologic diagnoses. CASE: A 22-year-old male presented with shoulder paresthesias and back pain, progressing to bilateral leg weakness. Spine MRI demonstrated an enhancing intramedullary T8 lesion, prompting urgent laminectomy and resection. Frozen section suggested a glial neoplasm. Four-month follow-up MRI spine showed leptomeningeal dissemination at the C-spine and caudal thecal sac. CSF analysis revealed 110 mg/dL protein, 2 cells/mcl, and negative cytology. NCI methylation profiling demonstrated 1p deletion and KIAA1549:BRAF fusion, consistent with DLGNT, subtype 1, with MAPK pathway activation; Ki-67 index was 30%. Treatment with binimetinib was initiated. After 1 month of therapy, the patient is tolerating treatment and strength is improving. DISCUSSION This case describes a rare presentation of a rare tumor. DLGNT, subtype 1, has a median age of diagnosis of 5 years and an indolent course. We describe an adult patient with molecular features consistent with DLGNT subtype 1, but a high proliferative index suggesting a more aggressive growth potential. In this case, methylation profiling aided in diagnosing the tumor type and guided treatment. The KIAA1549:BRAF fusion is an activating mutation in the MAPK pathway, and binimetinib was selected as a MEK inhibitor with high CNS penetration. To our knowledge, this is the first case of an adult-onset DLGNT treated with binimetinib. Based on our single patient experience, we would advocate for methylation profiling for rare CNS tumor entities.

Metastasis of a Primary Sinonasal Alveolar Rhabdomyosarcoma to the Testicle in an Adult Patient

Abstract Introduction/Objective Alveolar rhabdomyosarcoma is an aggressive tumor of skeletal muscle differentiation that predominantly affects adolescents and young adults, and is rare over 45 years of age. It has poor prognosis with metastasis, and most commonly spreads to the lymph nodes, bone marrow, and lungs. In this report, we describe a middle-aged patient with a primary sinonasal alveolar rhabdomyosarcoma which metastasized to the right testicle, an unusual metastatic site reported only a few times in literature. Methods/Case Report Our patient is a 53 year old male who presented in 2020 with recurrent epistaxis and bilateral cervical lymphadenopathy over a 2 month period. Maxillofacial CT/MRI imaging revealed a 4-cm sized mass within the left nasal cavity eroding into the ethmoid sinuses. The patient underwent endoscopic sinus surgery of the sinonasal mass, which on histology, appeared as poorly differentiated small round blue cells with numerous mitoses infiltrating as single cells and nests. These cells stained positive for vimentin, myogenin, desmin, and CD56, and negative for AE1/AE3, chromogranin, S-100, CD99, and CD45. The cells showed very high Ki67 proliferation index (70-80%) and tested positive for FOXO1 gene rearrangement, which supported the diagnosis of alveolar rhabdomyosarcoma. The patient completed chemoradiation therapy, and MRI scans over the next 2 years were negative for residual sinonasal tumor. However, the patient returned in 2022 after having undergone a right orchiectomy at an outside hospital for testicular pain, and abdominal CT showed diffuse retroperitoneal lymphadenopathy concerning for metastasis. On histology, the resected testicle showed seminiferous tubules infiltrated and destroyed by sheets of poorly differentiated small round blue cells with fibrous septae separating the nests of tumor cells into alveolar-like spaces with central loss of cohesion. The tumor cells showed the same immunohistochemical staining pattern as the primary sinonasal lesion, thus confirming it to be metastatic alveolar rhabdomyosarcoma. Results (if a Case Study enter NA) NA. Conclusion It is unusual for alveolar rhabdomyosarcomas to occur in adults over 45 years old, and is even rarer for these tumors to metastasize to the testicle. Metastases also only account for 1% of testicular tumors. Our report documents a very rare case of an alveolar rhabdomyosarcoma in a middle-aged adult which metastasized to the testicle after chemoradiation therapy.

Sarcomatoid Eccrine Porocarcinoma in a Patient with Urothelial Carcinoma

Abstract Introduction/Objective Sarcomatoid porocarcinoma of skin is an exceedingly rare and diagnostically challenging malignancy of the sweat gland. In this report, we describe a rare case of a sarcomatoid eccrine porocarcinoma initially diagnosed to be a squamous cell carcinoma (SCC) in a patient with metastatic urothelial carcinoma (UC). Methods/Case Report Our patient is a 60 year old male with stage IV, poorly differentiated UC status post TURBT with residual tumor, and a 4 x 3 x 1 cm, polypoid, ulcerating skin nodule on his right cheek presumptively diagnosed to be an SCC on biopsy at an outside hospital. Given the concern that this cutaneous lesion may be a metastatic UC with squamous differentiation, the skin nodule was re-biopsied at our hospital. On histology, the lesion consisted of sheets of squamoid, polygonal cells in the dermis with pleomorphic nuclei showing irregular contours, prominent nucleoli, coarse chromatin, and frequent mitoses. The tumor was focally connected to the epidermis, which established it as a primary skin neoplasm and not a metastatic lesion. The tumor cells stained diffusely positive for vimentin, AE1/AE3, CK5/6, HMW CK, CK7, p63, and p16, negative for CK20, GATA-3, BerEP4, and Sox10, and showed high Ki67 proliferation index. Within the tumor sheets were poorly formed, focal ductal elements that stained positive for CAM5.2, EMA, CEA, and CK19, and negative for p63. The tumor showed comedo necrosis, epidermal ulceration, and no keratinization. The tumor was diagnosed to be a porocarcinoma given its positive staining for CK7 and p16, presence of focal ductal elements, and epidermal connection, with sarcomatoid features indicated by strong vimentin expression. Results (if a Case Study enter NA) NA. Conclusion In this report, we described an exceedingly rare, diagnostically challenging case of a sarcomatoid porocarcinoma initially suspected to be a cutaneous SCC or metastatic UC with squamous differentiation. Although the tumor cells morphologically resembled SCC, its positive staining for CK7 and p16, presence of ductal elements, and lack of keratinization, favored a diagnosis of porocarcinoma rather than cutaneous SCC. Its focal connection to the epidermis and lack of GATA-3 expression favored a primary skin neoplasm rather than a metastatic UC. Its sarcomatoid nature was indicated by strong vimentin expression. Sarcomatoid porocarcinoma is diagnostically challenging given its rarity and potential to mimic other cutaneous malignancies such as SCC.

ODP020 A Challenging Case of Poorly Differentiated Carcinoma with Neuroendocrine Differentiation and Adrenal Mass

Abstract Background Neuroendocrine tumors (NETs) account for about 0.5% of all newly diagnosed malignancies. The most frequent primary sites are the gastrointestinal tract or pancreatic (62%-67%) and the lung (22%-27%). All poorly differentiated NETs have a Ki-67 index of greater than 20% and WHO grade of 3. The determination of the site of origin can be challenging in widely metastatic poorly differentiated NETs. Clinical Case A 51-year man with a history of hypertension and type 2 diabetes mellitus presented with angioedema while on lisinopril for years. Apart from a month's history of bilateral flank pain, he denied diarrhea, flushing, stridor, or shortness of breath. CT abdomen showed an 8.2×8. 0×7.9 cm right adrenal mass with a few punctate calcifications, hepatomegaly with multiple liver metastases, multiple retroperitoneal, hepatoduodenal, retrocrural adenopathy, and T4 metastatic lesion. CT chest showed bulky mediastinal and supraclavicular lymph nodes with trachea narrowing. All those lesions were FDG avid on PET/CT scan. Core biopsy of right supraclavicular lymph node revealed a poorly differentiated carcinoma with neuroendocrine differentiation and high proliferative index (25-50%). On cell block, it was negative for (AE1/AE3 (-), CAM5.2 (-), CK7, CK20, vimentin, desmin, CEA, EMA, CD45, CD20, and TTF-1 but positive for CK903 (focal +), synaptophysin (+), chromogranin (+), calretinin (focally +), inhibin (focally +). Based on the immunohistochemical stain, it was concluded as a metastatic small cell carcinoma of unknown origin. He was treated with carboplatin, etoposide, and atezolizumab and received 5 cycles. Endocrine was consulted two months after his initial presentation. Urine normetanephrine was &amp;gt; 4-fold elevated. Blood pressure was controlled on Doxazosin at 4 mg bid. The 10 gene-panel for hereditary pheochromocytoma and Paraganglioma (PGL) was negative. There was a progression of mediastinal adenopathy while on treatment with narrowing of proximal intrathoracic trachea requiring elective tracheostomy. Based on imaging findings and elevated normetanephrine, we believe he has metastatic pheochromocytoma and PGL. His chemo regimen was switched to Cyclophosphamide, Vincristine, and Dacarbazine (CVD). He finished 8 cycles of CVD with an overall reduction in the size of lymph nodes (both above and below the diaphragm) and right adrenal mass. His plasma normetanephrine level was also significantly improved. Presentation: No date and time listed

Capecitabine and temozolomide for metastatic intermediate to high-grade pancreatic neuroendocrine neoplasm: a single center experience

The combination of capecitabine and temozolomide (CAPTEM) is one of the treatment options for metastatic pancreatic neuroendocrine neoplasms (pNENs). This study aims to evaluate the efficacy of CAPTEM in patients with metastatic intermediate to high-grade pancreatic neuroendocrine tumor (pNET) or carcinoma (pNEC). This study was conducted retrospectively in a single center. Patients were treated for intermediate to high-grade tumor with 750 mg/m² of capecitabine twice daily from day 1 to 14 and 200 mg/m² of temozolomide once daily from day 10 to 14, repeating twice in a cycle of 28 days. The primary outcomes were durations of overall survival (OS) and progression-free survival (PFS). The secondary outcomes consisted of objective response rate and disease control rate. A total of 12 patients (grade 2 NET in six, grade 3 NET in three, NEC in three patients) who received CAPTEM were included in this study. Patients received a median of five cycles (range, 2 to 46) of CAPTEM. The median dose combined 1,150 mg of capecitabine and 300 mg of temozolomide. The median OS and PFS were 41.2 months (range, 3.2 to 167) and 39.7 months (range, 2.1 to 100), respectively. Patients with NET had longer OS and PFS compared to those of patients with NEC (p = 0.002 and p = 0.028). High Ki-67 proliferative index (&gt; 50%) was significantly associated with poor survival outcomes. CAPTEM showed favorable survival outcomes in patients with metastatic intermediate to high-grade pNENs. Our study supports that CAPTEM may be an effective treatment option for metastatic pNENs.

Pulmonary papillary adenoma with malignant potential: a case report and literature review

BackgroundPulmonary papillary adenoma is a rare benign tumor in the periphery of the lung. We report a 66-year-old female patient with a tumor in the lower lobe of the right lung and present the clinicopathological features and review the literature.Case presentationA tumor in the lower lobe of the right lung was found incidentally on chest X-ray during the physical examination of the patient, and the patient occasionally had a dry cough that was not treated. The tumor was clearly demarcated and lobulated on CT scan. After 2 years of follow-up, the boundary of the tumor was still clear, with more lobulations and the enhanced scan showed uniform enhancement. Grossly, the tumor had a granular cut surface and was easy to fall off, which was helpful for the diagnosis of papillary adenoma during intraoperative frozen examination. Under the microscope, most areas of the tumor had the typical morphological structure of papillary adenoma. However, the tumor locally protruded into the surrounding lung tissue, accompanied by crowded cells and high cell proliferation index. It was suggested that this case of papillary adenoma had malignant potential and needed active intervention and treatment.ConclusionPulmonary papillary adenoma is a rare epithelial tumor with malignant potential. Surgical treatment should be performed as soon as possible after diagnosis to prevent malignant transformation.

CD47 expression and tumor-associated immune cells in breast cancer and their correlation with molecular subtypes and prognostic factors.

Breast cancer is the most common malignancy in women and a heterogeneous disease at the molecular level. Since most breast cancer cases are not of a special type, it is suggested that tumor-associated macrophages and tumor-infiltrating lymphocytes, which are involved in tumor growth, invasion, angiogenesis, and metastasis, may be important factors that should be evaluated together with standard criteria to determine the prognosis of cancer and assist in treatment decisions and outcome stratification. In this study, CD47 expression, which is involved in macrophage-mediated immune escape, tumor-infiltrating lymphocytes, and tumor-associated macrophages were evaluated in breast cancer molecular subgroups and correlated with prognostic factors. The immunohistochemistry of CD47, CD163, and CD3 was analyzed on the tissue microarrays of 278 invasive breast cancer cases. The CD47, CD163, and CD3 expressions were found to be correlated with various clinicopathological parameters in breast cancer. High levels of CD47, CD163, and CD3 expressions had a significant correlation with the ER status and PR status, Ki-67 proliferation index, and molecular subtype (P<0.05). The CD47 expression had a significant correlation with the CD3 and CD163 expressions (p=0.021 and p=0.001, respectively). Our results suggest that CD47, CD163, and CD3 may be among the prognostic factors of breast cancer. The combined use of CD47, CD163, and CD3 can be a new prognostic factor for patients with breast cancer, especially as a therapeutic target in hormone receptor-negative breast cancer cases and those with a high proliferation index.

STAT-5 and STAT-6 in Breast Cancer: Potential Crosstalk With Estrogen and Progesterone Receptors Can Affect Cell Proliferation and Metastasis.

Signal transducers and activators of transcription 5a and 6 (STAT5a and STAT6) play a critical role in tumorigenesis of mammary glands. Based on previous studies, the breast cancer is largely dependent on hormone receptors. Consequently, it is very interesting to decipher the relationship between the STAT5a and STAT6 expression and the molecular distribution of estrogen receptors (ERs) and progesterone receptors (PRs) in mammary tumors. Our study analyzed the expression of STAT5a and STAT6, ERα, ERβ and PR in 40 breast tumor tissues using quantitative real-time polymerase chain reaction (qRT-PCR). Furthermore, the Ki-67 and HER2 status were detected using immunohistochemistry. STAT5a and STAT6 were retained in the majority of the cases studied. Increasing of STAT5a and STAT6 is significantly associated with ERs and PR. The coexpression of both STAT5a and STAT6 with ERs and PR is associated with high tumor grades. Moreover, the coexpression of STAT5a and STAT6 with ERα and PR is associated with a high proliferation index. In addition, (STAT6 + ERβ+) and (STAT6 + PR+) breast cancer subgroups are associated with lymph node infiltration (P = 0.001 and P = 0.03, respectively). Our study results provide an interaction between STAT5a and STAT6 with ERs and PR inducing cell proliferation. Coexpression of STAT5a and STAT6 with ERs and PR can predict sensibility to hormonal therapy.