Novel targetable biomarkers in clear cell carcinoma of the breast uncovered by molecular profiling: A study of nine cases.

We profiled nine pure clear cell carcinomas of the breast using massively parallel DNA and RNA sequencing (NGS), in situ hybridization (ISH), and immunohistochemistry (IHC). All cases were primary mammary clear cell carcinomas that were diagnosed in female patients (mean age: 53.4 years; range: 31-69 years). Based on our findings, we conclude that the majority of clear cell carcinomas are ER/PR positive and consequently amenable to anti-ER treatment modalities. A subset of clear cell carcinomas also harbored alterations in PIK3CA/PTEN/AKT pathway, particularly PTEN, indicating a potential benefit of PI3K/Akt/mTOR inhibitors. The status of I-O biomarkers in clear cell carcinomas indicates a limited therapeutic benefit of immune checkpoint inhibitors (against PD-1/PD-L1)

EL AGROTURISMO COMO FUENTE DE EMPRENDIMIENTO COMUNITARIO. CASO DE ESTUDIO: : LA COLOMBIA ALTA.

Rural tourism integrates nature, culture and the active participation of locals, whose objective is to improve the living conditions of the community (Baroroh, Wahjoedi, Utomo and Lestari, 2021). However, no studies have been found to analyze the potential of agrotourism in the La Colombia Alta site, located in the province of Bolívar, Ecuador. Therefore, the purpose of this case study is to analyze the empirical practices of the main promoter in that locality and to evaluate the impact of agrotourism in the sector; by studying and describing the perception and degree of satisfaction of tourists to identify the priority aspects to improve. This study is mixed, under a descriptive, longitudinal approach. Interviews, surveys and field visits were the main collection instruments. The findings proved that 1. the profile of the main entrepreneurial promoter in the sector coincides with those of other studies. 2. the perception of visitors was positive (91.4%) although the level of satisfaction differed. 3. the level of impact of agrotourism in the sector has a positive incidence (80% of tourists estimate to return and incurrence to date of 3.14 times). Therefore, it is recommended to carry out the continuity of the case, after the application of change actions to improvements based on the suggestions given in this article. The results could have implications in the research field of local agrotourism and its real application for the improvement of the needs of the communities.El turismo rural integra naturaleza, cultura y participación activa de los locales, cuyo objetivo es mejorar la condición de vida de la comunidad al aprovechar los flujos económicos que el mismo produce. Sin embargo, no se han encontrado estudios para analizar el potencial del agroturismo en el recinto La Colombia Alta, ubicada en la provincia de Bolívar, Ecuador.   Por lo tanto, el presente caso de estudio tuvo como propósito analizar las prácticas empíricas de la principal promotora en dicha localidad y evaluar el impacto del agroturismo en el sector; al estudiar y describir la percepción y grado de satisfacción de los turistas para identificar los aspectos prioritarios a mejorar. Este estudio es mixto, bajo un enfoque descriptivo, de corte longitudinal, realizado a través de entrevistas, encuestas y visitas de campos siendo las principales técnicas e instrumentos de recolección de datos. Los hallazgos probaron que 1. el perfil de la principal promotora emprendedora del sector coincide con los de otros estudios. 2. la percepción de los visitantes fue positiva (91,4%) y nivel de satisfacción difiere 3. el nivel de impacto del agroturismo en el sector tiene una incidencia positiva (80% de los turistas estiman regresar e incurrencia a la fecha de 3.14 veces). Por consiguiente, se recomienda llevar la continuidad del caso, tras la aplicación de acciones de cambio a mejoras en base a las sugerencias dadas en este artículo. Los resultados podrían tener implicaciones en el campo investigativo del agroturismo local y a su aplicación real para el mejoramiento de las necesidades de las comunidades

Comparison of the biomarkers for targeted therapies in primary extra-mammary and mammary Paget's disease.

Primary Extra-mammary Paget's disease (EMPD) is a very rare cutaneous adenocarcinoma affecting anogenital or axillary regions. It is characterized by a prolonged course with recurrences and eventually distant metastatic spread for which no specific therapy is known. Eighteen EMPD (13 vulvar and five scrotal) and ten mammary Paget's disease (MPD) cases were comprehensively profiled for gene mutations, fusions and copy number alterations, and for therapy-relevant protein biomarkers). Mutations in TP53 and PIK3CA were the most frequent in both cohorts: 7/15 and 5/15 in EMPD; 1/6 and 4/7 in MPD HER2 gene amplification was detected in 4/18 EMPD (3 vulvar and 1 scrotal case) in contrast to MPD where it was detected in the majority (7/8) of cases. TOP2A gene amplification was seen in 2/12 EMPD and 1/6 MPD, respectively. Similarly, no difference in estrogen receptor expression was seen between the EMPD (4/15) and MPD (3/10). Androgen receptor was also expressed in the majority of both cohorts (12/16 EMPD) and (7/8 MPD).Here ARv7 splice variant was detected in 1/7 EMPD and 1/4 MPD cases, respectively. PD-L1 expression on immune cells was exclusively observed in three vulvar EMPD. In contrast to MPD, six EMPDs harbored a "high" tumor mutation burden (≥10 mutations/Mb). All tested cases from both cohorts were MSI stable. EMPD shares some targetable biomarkers with its mammary counterpart (steroid receptors, PIK3CA signaling pathways, TOP2A amplification). HER2 positivity is notably lower in EMPD while biomarkers to immune checkpoint inhibitors (high TMB and PD-L1) were observed in some EMPD. Given that no consistent molecular alteration characterizes EMPD, comprehensive theranostic profiling is required to identify individual patients with targetable molecular alterations

Potential Novel Therapy Targets in Neuroendocrine Carcinomas of the Breast.

Neuroendocrine carcinoma (NEC) of the breast is a rare, special type of breast cancer, reportedly constituting 2% to 5% of all breast cancers. Although breast NEC does not have a specific targeted therapy, several new targeted therapies based on specific biomarkers were recently investigated in the NEC of lung and in other types of breast carcinoma, which may provide guidance to their feasibility in breast NEC. Twenty breast NECs were profiled for biomarkers of therapy including antibody-drug conjugates (DLL3, TROP-2, and FOLR1), histone deacetylase (H3K36Me3) inhibitors, tropomyosin receptor kinases (NTRK1/2/3 gene fusions) targeted inhibitors, alkylating agents (MGMT), and immune checkpoint inhibitors (PD-L1, TMB, and MSI) using immunohistochemistry and DNA/RNA next-generation sequencing assays. Predictive expression of TROP-2, FOLR1, and H3K36Me3 were detected in different subsets of tumors and may pave the way for development of novel targeted therapies in some patients with breast NECs. There was no evidence of DLL3 expression, NTRK gene fusions, or MGMT hypermethylation. No biomarkers predictive of immune checkpoint inhibitor efficacy (programmed death-ligand 1 expression, tumor mutational burden, microsatellite instability) were identified. FGFR and CCND1 gene amplifications were detected in isolated cases. This study identified several potential targets for novel therapies in breast NEC, including farletuzumab and mirvetuximab soravtansine (FOLR1), sacituzumab govitecan (TROP-2), and HDAC inhibitors (H3K36Me3). In some cases, CCND1 gene amplification may indicate the usefulness of investigational therapies. The reported results should serve as an early indication of potential clinical relevance in selected patients with breast NEC

TERT gene fusions characterize a subset of metastatic Leydig cell tumors

Objective: Metastatic Leydig cell tumors (LCT) are rare, difficult to treat malignancies without known underlying molecular-genetic events. An index case of metastatic LCT showed an LDLR-TERT gene fusion upon routine genetic profiling for detection of therapeutic targets, which was then followed by an investigation into a cohort of additional LCTs. Patients and Methods: Twenty-nine LCT (27 male and 2 female patients) were profiled using NGS and immunohistochemistry. Results: TERT gene fusions were detected only in testicular metastatic Leydig cell tumors, in three of seven successfully analyzed cases (RMST:TERT, LDLR:TERT and B4GALT5:TERT). TOP1 and CCND3 amplifications were identified in the case with a B4GALT5:TERT fusion. A TP53 mutation was detected in one metastatic tumor without a TERT fusion. Five primary (four testicular and one ovarian) LCTs showed multiple gene amplifications, without a consistent pattern. A single metastatic ovarian LCT showed BAP1 mutation and copy number amplifications affecting the NPM1, PCM1 and SS18 genes. At the protein level, 4/7 metastatic and 6/10 primary testicular LCTs over-expressed TOP1. Androgen receptor (AR) was overexpressed in 10/13 primary testicular tumors and 2/5 metastatic testicular LCT (without detectable ARv7 mRNA or ARv7 protein). Only one metastatic testicular LCT exhibited high TMB while all tested cases were MSI stable and did not express PD-L1. Conclusions: Our study for the first time identified TERT gene fusions as a main genetic alteration and a potential therapeutic target in metastatic Leydig cell tumors. TOP1 and AR may guide decisions on chemo- and/or hormone therapy for selected individual patients.Qatar National Librar

Survey Data on the Impact of COVID-19 on Parental Engagement Across 23 Countries

This data article describes the dataset of the International COVID-19 Impact on Parental Engagement Study (ICIPES). ICIPES is a collaborative effort of more than 20 institutions to investigate the ways in which, parents and caregivers built capacity engaged with children's learning during the period of social distancing arising from global COVID-19 pandemic. A series of data were collected using an online survey conducted in 23 countries and had a total sample of 4,658 parents/caregivers. The description of the data contained in this article is divided into two main parts. The first part is a descriptive analysis of all the items included in the survey and was performed using tables and figures. The second part refers to the construction of scales. Three scales were constructed and included in the dataset: ‘parental acceptance and confidence in the use of technology’, ‘parental engagement in children's learning’ and ‘socioeconomic status’. The scales were created using Confirmatory Factor Analysis (CFA) and Multi-Group Confirmatory Analysis (MG-CFA) and were adopted to evaluate their cross-cultural comparability (i.e., measurement invariance) across countries and within sub-groups. This dataset will be relevant for researchers in different fields, particularly for those interested in international comparative education

Survey data on the impact of COVID-19 on parental engagement across 23 countries

This data article describes the dataset of the International COVID-19 Impact on Parental Engagement Study (ICIPES). ICIPES is a collaborative effort of more than 20 institutions to investigate the ways in which, parents and caregivers built capacity engaged with children's learning during the period of social distancing arising from global COVID-19 pandemic. A series of data were collected using an online survey conducted in 23 countries and had a total sample of 4,658 parents/caregivers. The description of the data contained in this article is divided into two main parts. The first part is a descriptive analysis of all the items included in the survey and was performed using tables and figures. The second part refers to the construction of scales. Three scales were constructed and included in the dataset: 'parental acceptance and confidence in the use of technology', 'parental engagement in children's learning' and 'socioeconomic status'. The scales were created using Confirmatory Factor Analysis (CFA) and Multi-Group Confirmatory Analysis (MG-CFA) and were adopted to evaluate their cross-cultural comparability (i.e., measurement invariance) across countries and within sub-groups. This dataset will be relevant for researchers in different fields, particularly for those interested in international comparative education

Molecular Profiling of Clear Cell Carcinoma of the Breast Reveals Novel Targetable Biomarkers

Background: Although clear cell morphology may be seen in various subtypes of breast cancer, a pure variant of clear cell carcinoma (usually glycogen-rich) is a very rare primary breast malignancy. It is characterized by the neoplastic cells with abundant and clear cytoplasm that contains glycogen. A recent SEER study highlighted the aggressive clinical behavior of this rare cancer. Apart from the steroid receptors profile (ER-positive in 33-75%, PR positive in 30-43% and HER2 positive in 0-44%) status, no information is available regarding its molecular features and targetable biomarkers. Design: Nine pure (>90% clear cell morphology) clear cell carcinomas of the breast were comprehensively profiled using massively parallel DNA and RNA sequencing (NGS), in-situ hybridization and immunohistochemistry. Results: Steroid receptors ER and AR were positive in 8/9 and 7/9 cases, respectively. AR was positive in 6/7 cases without the presence of ARv7 splice variant. None of the cases was HER2 positive. Pathogenic mutations were found in PIK3R1 and BRCA2 (#1), TP53, PTEN and CDKN2A (#2), TP53 and BCOR1 (#3). PTEN protein loss was confirmed by IHC in PTEN mutated cases as well as in two additional cases without detectable PTEN gene mutation. No gene fusion was seen in any of the cases. Low PD-L1 expression (1-10%) was exclusively seen in immune cells (n=3/8). All tested cases (n=8) were MSI stable and had low TMB (3-7 mutations/mb) (n=3). Conclusion: Clear cell carcinomas are predominantly ER and AR-positive and consequently amenable for endocrine treatment regimes. Frequent AR over-expression without ARv7 may support trials with anti-AR therapies. A proportion of clear cell carcinomas harbors alterations in the PIK3CA/PTEN pathway indicating a potential benefit of PI3K/Akt/mTOR inhibitors while the presence of PD-L1 on immune cells warrants the trials with immune checkpoint inhibitors.Caris Life Sciences, Phoenix, Arizona, US

Abstract B094: Novel method for patient stratification in breast carcinoma based upon spatial analysis of tumor microenvironment

Breast cancer consists of several intrinsic molecular subtypes, providing the basis for clinical treatment decisions. Lately, it is becoming increasingly recognized that factors other than the intrinsic cancer characteristics, such as immune components’ activity in the tumor microenvironment, have important effects on treatment choices and efficacy. bioSyntagma has developed a method, the Molecular Fingerprint (mPrint®), that enables multiplexed analysis of spatially defined regions in formalin-fixed, paraffin-embedded (FFPE) tumor samples allowing for analysis of the gene signatures unique to the tumor microenvironment. This method was applied to molecularly defined sets of breast cancers and used to evaluate four different tumor regions of interest (ROIs): 1) viable carcinoma proper (>90% cancer cells), 2) fibrotic tumor center (sparse cellularity), 3) interface between viable tumor and inflammatory component (tumor and inflammatory microenvironment) and 4) tissue away from the tumor (normal breast tissue). This was compared to the whole tissue scrapes from each patient block. Each ROI and tissue scrape was analyzed by high throughput qPCR for a panel of 248 genes using SmartChip technology (Takara Bio, USA). Sequential tissue slices from each patient were also analyzed using immunohistochemistry (IHC) for three targets and investigated for correlation with qPCR results for validation of the method. Overall, reasonable concordance was observed in general expression trends between selected IHC and RNA expression. qPCR data were further analyzed using hierarchical clustering analysis and showed that morphologically defined ROI’s cluster completely differently than traditional clustering of entire tissue scrapes. Notably, patient clustering based on morphological regions was independent of the intrinsic cancer subtype, as determined by molecular profiling of whole tissue scrapes, as well as independent of trends in Tumor Mutational Burden (TMB) and Microsatellite Instability (MSI). These findings suggest that current methods of patient stratification based on whole tumor molecular subtyping may be inferior to stratification based on molecular characteristics of the tumor microenvironment.bioSyntagma, LLC, PHOENIX, AZ; Takara Bio USA, Inc., Mountain View, CA

Comparison of the biomarkers for targeted therapies in primary extra‐mammary and mammary Paget's disease

Abstract Background Primary Extra‐mammary Paget's disease (EMPD) is a very rare cutaneous adenocarcinoma affecting anogenital or axillary regions. It is characterized by a prolonged course with recurrences and eventually distant metastatic spread for which no specific therapy is known. Methods Eighteen EMPD (13 vulvar and five scrotal) and ten mammary Paget's disease (MPD) cases were comprehensively profiled for gene mutations, fusions and copy number alterations, and for therapy‐relevant protein biomarkers). Results Mutations in TP53 and PIK3CA were the most frequent in both cohorts: 7/15 and 5/15 in EMPD; 1/6 and 4/7 in MPD HER2 gene amplification was detected in 4/18 EMPD (3 vulvar and 1 scrotal case) in contrast to MPD where it was detected in the majority (7/8) of cases. TOP2A gene amplification was seen in 2/12 EMPD and 1/6 MPD, respectively. Similarly, no difference in estrogen receptor expression was seen between the EMPD (4/15) and MPD (3/10). Androgen receptor was also expressed in the majority of both cohorts (12/16 EMPD) and (7/8 MPD).Here ARv7 splice variant was detected in 1/7 EMPD and 1/4 MPD cases, respectively. PD‐L1 expression on immune cells was exclusively observed in three vulvar EMPD. In contrast to MPD, six EMPDs harbored a “high” tumor mutation burden (≥10 mutations/Mb). All tested cases from both cohorts were MSI stable. Conclusions EMPD shares some targetable biomarkers with its mammary counterpart (steroid receptors, PIK3CA signaling pathways, TOP2A amplification). HER2 positivity is notably lower in EMPD while biomarkers to immune checkpoint inhibitors (high TMB and PD‐L1) were observed in some EMPD. Given that no consistent molecular alteration characterizes EMPD, comprehensive theranostic profiling is required to identify individual patients with targetable molecular alterations