Distinct Responsiveness Research Articles

Mediastinal germ cell tumors: a narrative review of their traits and aggressiveness features.

Mediastinal extragonadal germ-cell tumors (MEGCTs) are rare neoplasms with a multifaceted clinical behavior. This paper is devoted to review their main characteristics, including histological patterns and different factors of aggressiveness in MEGCTs. Proper understanding of the latter can help to better stratify patients' prognoses and improve clinical management. Different theories exist on the origin of MEGCTs, including primordial germ cells deposition during embryogenesis. MEGCTs predominantly affects young males and aggressiveness follows the ability of local and systemic spread of each germ-cell neoplasia subtype, as well as their distinct responsiveness to therapy. Indeed, non-seminomatous MEGCTs have a worse prognosis. Unfortunately, they are also more frequent than seminomas in the mediastinum. Regardless of histological type, local aggressiveness can follow tumoral expansion with compression on or infiltration of mediastinal structures. Chemotherapy can be effective in reducing neoplastic volume, but different levels of sensitivity can be found in different MGCTs. In particular, a chemo-resistant teratoma component of a mixed MEGCTs can undergo a paradoxical enlargement after chemotherapy, while other components of the tumor regress. This is reflected by a concomitant normalization of serum tumoral markers and cardiopulmonary deterioration due to compression. Such clinical phenomenon, called growing-teratoma syndrome (GTS), requires a prompt surgical approach. A literature research of pertinent epidemiological, pathological and clinical articles was conducted. The mediastinum can harbor different kinds of neoplasia, including GCTs. The full spectrum of MEGCTs includes a variety of tumors with different clinical behaviors. Aggressiveness follows the inherent ability of local and systemic spread of each neoplastic type, as well as their distinct responsiveness to therapy.

Lung Microbiota Signature and Corticosteroid Responses in Pneumonia-Associated Acute Respiratory Distress Syndrome in Hematological Patients.

ObjectiveIn this study, we aim to classify hematological patients with the pneumonia-associated acute respiratory syndrome (ARDS) into different groups that were characterized by distinct early responsiveness to corticosteroids, describe the microbiota signatures of the non-responders and responders, and compare the prognosis of the two groups.MethodsHematological patients with ARDS were included and treated with mechanical ventilation and corticosteroid. According to the early improvement to the corticosteroid therapy, patients were classified as non-responders and responders. The lung microbiota signatures and the outcomes of the non-responders and responders were compared.ResultsFifty patients were included in this study. Twenty-eight patients were classed as non-responders and 22 as responders. Compared to the non-responders, responders had higher serum levels of IL-6, IL-8, TNF-α and CRP, their lung microbiota was with lower alpha diversity and enriched with virus species. The responders had an overall higher ventilator free days than the non-responders [4 (0–6) vs 6 (0–10), p=0.034], for survivors the difference was more significant [5 (3–6) vs 8 (3–10), p=0.012]. Survival analysis showed that there was no difference in survival rate between the two groups over time (Log-rank p=0.073). When non-responders were stratified into subgroups of patients with infection or co-infection, those non-responders with co-infection had significantly lower survival rate than other patients (Log-rank p= 0.028).ConclusionFor hematological patients with pneumonia-associated ARDS, the responders of corticosteroids had higher ventilator free days at day 28 than the non-responders. The microbiota signatures were distinct in the two groups. The non-responders with coinfections had the lowest survival rate when compared to the non-responders with no coinfections and the responders.

Identification and validation of a novel signature for prediction the prognosis and immunotherapy benefit in bladder cancer.

BackgroundBladder cancer (BC) is a common urinary tract system tumor with high recurrence rate and different populations show distinct response to immunotherapy. Novel biomarkers that can accurately predict prognosis and therapeutic responses are urgently needed. Here, we aim to identify a novel prognostic and therapeutic responses immune-related gene signature of BC through a comprehensive bioinformatics analysis.MethodsThe robust rank aggregation was conducted to integrate differently expressed genes (DEGs) in datasets of the Cancer Genome Atlas (TCGA) and the gene expression omnibus (GEO). Lasso and Cox regression analyses were performed to formulate a novel mRNA signature that could predict prognosis of BC patients. Subsequently, the prognostic value and predictive value of the signature was validated with two independent cohorts GSE13507 and IMvigor210. Finally, quantitative Real-time PCR (qRT-PCR) analysis was conducted to determine the expression of mRNAs in BC cell lines (UM-UC-3, EJ-1, SW780 and T24).ResultsWe built a signature comprised the eight mRNAs: CNKSR1, COPZ2, CXorf57, FASN, PCOLCE2, RGS1, SPINT1 and TPST1. Our prognostic signature could be used to stratify BC population into two risk groups with distinct immune profile and responsiveness to immunotherapy. The results of qRT-PCR demonstrated that the eight mRNAs exhibited different expression levels in BC cell lines.ConclusionOur study constructed a convenient and reliable 8-mRNA gene signature, which might provide prognostic prediction and aid treatment decision making of BC patients in clinical practice.

Immunological Classification of Pancreatic Carcinomas to Identify Immune Index and Provide a Strategy for Patient Stratification.

BackgroundCancer immunotherapy has produced significant positive clinical effects in a variety of tumor types. However, pancreatic ductal adenocarcinoma (PDAC) is widely considered to be a “cold” cancer with poor immunogenicity. Our aim is to determine the detailed immune features of PDAC to seek new treatment strategies.MethodsThe immune cell abundance of PDAC patients was evaluated with the single-sample gene set enrichment analysis (ssGSEA) using 119 immune gene signatures. Based on these data, patients were classified into different immune subtypes (ISs) according to immune gene signatures. We analyzed their response patterns to immunotherapy in the datasets, then established an immune index to reflect the different degrees of immune infiltration through linear discriminant analysis (LDA). Finally, potential prognostic markers associated with the immune index were identified based on weighted correlation network analysis (WGCNA) that was functionally validated in vitro.ResultsThree ISs were identified in PDAC, of which IS3 had the best prognosis across all three cohorts. The different expressions of immune profiles among the three ISs indicated a distinct responsiveness to immunotherapies in PDAC subtypes. By calculating the immune index, we found that the IS3 represented higher immune infiltration, while IS1 represented lower immune infiltration. Among the investigated signatures, we identified ZNF185, FANCG, and CSTF2 as risk factors associated with immune index that could potentially facilitate diagnosis and could be therapeutic target markers in PDAC patients.ConclusionsOur findings identified immunologic subtypes of PDAC with distinct prognostic implications, which allowed us to establish an immune index to represent the immune infiltration in each subtype. These results show the importance of continuing investigation of immunotherapy and will allow clinical workers to personalized treatment more effectively in PDAC patients.

Raspberry consumption: identification of distinct immune-metabolic response profiles by whole blood transcriptome profiling

Numerous studies have reported that diets rich in phenolic compounds are beneficial to immune-metabolic health, yet these effects are heterogeneous and the underlying mechanisms are poorly understood. To investigate the inter-individual variability of the immune-metabolic response to raspberry consumption, whole-blood RNAseq data from 24 participants receiving 280 g/d of raspberries for 8 weeks were used for the identification of responsiveness subgroups by using partial least squares-discriminant analysis (PLSDA) and hierarchical clustering. Transcriptomic-based clustering regrouped participants into two distinct subgroups of 13 and 11 participants, so-called responders and non-responders, respectively. Following raspberry consumption, a significant decrease in triglycerides, cholesterol and C-reactive protein levels were found in responders, as compared to non-responders. Two major gene expression components of 100 and 220 genes were identified by sparse PLSDA as those better discriminating responders from non-responders, and functional analysis identified pathways related to cytokine production, leukocyte activation and immune response as significantly enriched with most discriminant genes. As compared to non-responders, the plasma lipidomic profile of responders was characterized by a significant decrease in triglycerides and an increase in phosphatidylcholines following raspberry consumption. Prior to the intervention, a distinct metagenomic profile was identified by PLSDA between responsiveness subgroups, and the Firmicutes-to-Bacteroidota ratio was found significantly lower in responders, as compared to non-responders. Findings point to this transcriptomic-based clustering approach as a suitable tool to identify distinct responsiveness subgroups to raspberry consumption. This approach represents a promising framework to tackle the issue of inter-individual variability in the understanding of the impact of foods on immune-metabolic health.

Cholinergic relevant functional reactivity is associated with dopamine responsiveness of tremor in Parkinson’s disease

Tremor in Parkinson’s disease (PD) has distinct responsiveness to dopamine, which is supposed not be exclusively related to dopamine deficiency but has a close relationship with cholinergic system. This phenomenon indicates that cholinergic system may be an important regulatory for distinct dopamine responsiveness of parkinsonian tremor. Through investigating the alterations of cholinergic and dopaminergic network during levodopa administration, we aimed at exploring the mechanisms of differed dopamine responsiveness of parkinsonian tremor. Fifty-two PD patients with tremor were enrolled. MRI scanning, UPDRS III and its sub-symptom scores were collected in OFF and ON status (dopaminergic challenge test). Then, patients were divided into two groups (dopamine-resistant tremor and dopamine-responsive tremor) according to the tremor change rate median score. Dopaminergic and cholinergic network were obtained. LASSO regression was conducted to identify functional connectivity with distinct reactivity during levodopa administration between groups. Afterwards, detailed group comparisons, interaction and correlation analyses were performed. The reactivity of cholinergic connectivity showed the highest possibility to distinguish two groups, especially connectivity of right basal forebrain 123 to right parietal operculum cortex (R.BF123-R.PO). After levodopa administration, connectivity of R.BF123-R.PO was decreased for dopamine-responsive tremor while which remained unchanged for dopamine-resistant tremor. The reactivity of R.BF123-R.PO was negatively correlated with tremor change rate. Reduced cholinergic connectivity to parietal operculum may be an underlying mechanism for the responsive tremor in PD and the distinct cholinergic reactivity of parietal operculum to levodopa may be a core pathophysiology for the differed DA responsiveness of tremor in PD.

The distinct responsiveness of cytokeratin 19-positive hepatocellular carcinoma to regorafenib

Cytokeratin 19-positive (CK19+) hepatocellular carcinoma (HCC) is an aggressive subtype characterized by early recurrence and chemotherapy tolerance. However, there is no specific therapeutic option for CK19+ HCC. The correlation between tumor recurrence and expression status of CK19 were studied in 206 patients undergoing liver transplantation for HCC. CK19−/+ HCC cells were isolated to screen effective antitumor drugs. The therapeutic effects of regorafenib were evaluated in patient-derived xenograft (PDX) models from 10 HCC patients. The mechanism of regorafenib on CK19+ HCC was investigated. CK19 positiveness indicated aggressiveness of tumor and higher recurrence risk of HCC after liver transplantation. The isolated CK19+ HCC cells had more aggressive behaviors than CK19− cells. Regorafenib preferentially increased the growth inhibition and apoptosis of CK19+ cells in vitro, whereas sorafenib, apatinib, and 5-fluorouracil did not. In PDX models from CK19−/+ HCC patients, the tumor control rate of regorafenib achieved 80% for CK19+ HCCs, whereas 0% for CK19− HCCs. RNA-sequencing revealed that CK19+ cells had elevated expression of mitochondrial ribosomal proteins, which are essential for mitochondrial function. Further experiments confirmed that regorafenib attenuated the mitochondrial respiratory capacity in CK19+ cells. However, the mitochondrial respiration in CK19− cells were faint and hardly repressed by regorafenib. The mitochondrial respiration was regulated by the phosphorylation of signal transducer and activator of transcription 3 (STAT3), which was inhibited by regorafenib in CK19+ cells. Hence, CK19 could be a potential marker of the therapeutic benefit of regorafenib, which facilitates the individualized therapy for HCC. STAT3/mitochondria axis determines the distinct response of CK19+ cells to regorafenib treatment.

Distinct hemispherical responsiveness to transcranial static magnetic field stimulation?

Distinct hemispherical responsiveness to transcranial static magnetic field stimulation?

Characterization of Molecular Subtypes in Head and Neck Squamous Cell Carcinoma With Distinct Prognosis and Treatment Responsiveness.

Head and neck squamous cell carcinoma (HNSCC) is one of the most aggressive malignancies with complex phenotypic, etiological, biological, and clinical heterogeneities. Previous studies have proposed different clinically relevant subtypes of HNSCC, but little is known about its corresponding prognosis or suitable treatment strategy. Here, we identified 101 core genes from three prognostic pathways, including mTORC1 signaling, unfold protein response, and UV response UP, in 124 pairs of tumor and matched normal tissues of HNSCC. Moreover, we identified three robust subtypes associated with distinct molecular characteristics and clinical outcomes using consensus clustering based on the gene expression profiles of 944 HNSCC patients from four independent datasets. We then integrated the genomic information of The Cancer Genome Atlas (TCGA) HNSCC cohort to comprehensively evaluate the molecular features of different subtypes and screen for potentially effective therapeutic agents. Cluster 1 had more arrested oncogenic signaling, the highest immune cell infiltration, the highest immunotherapy and chemotherapeutic responsiveness, and the best prognosis. By contrast, Cluster 3 showed more activated oncogenic signaling, the lowest immune cell infiltration, the lowest immunotherapy and chemotherapy responsiveness, and the worst prognosis. Our findings corroborate the molecular diversity of HNSCC tumors and provide a novel classification strategy that may guide for prognosis and treatment allocation.

Bulk density and porosity of soils influenced by skidding operations in the Nkrankwanta Off-Forest Reserve of Ghana

Heavy forest machinery used in skidding has the capacity to influence the physical properties of soils. These may possibly lead to an upsurge in soil disruption and compaction of the soil surface decreases forest soil fertilities. This study assesses the effects of skidding on some soil physical properties such as the soil bulk density and porosity in the Nkrankwanta off-forest reserve in Ghana. The treatments comprised of four traffic intensity levels (1, 5, 10, and 15 passes) of the Mercedes Benz skidder (MB) Trac 1800 and a slope of two levels (less than 20 % and greater than 20 %) in a completely randomized block design. In addition, porosity and soil bulk density were assessed at varied distances from the MB Trac 1800. Soil bulk density results showed increasing trends with traffic frequency. Soil bulk density measured in the undisturbed area was 0.64 g cm-3 and 0.56 g cm-3 at slopes of less than 20% and greater than 20%, respectively. On the skid trail, soil bulk density significantly increased with traffic frequency (p<0.05). However soil porosity declined. Soil porosity estimated in uninterrupted area was 59.10 % and 57.40 % at < 20% and > 20% slope, respectively. Soil porosity was significantly influenced via different skidder passes (p<0.05). The soil physical properties were not influenced by the steepness of the slope however acted together in the number of passes to influence soil porosity. The impacts of the skidder on soil physical properties were significantly apparent at distances of 2 m to each sideway of the skidding trail. In conclusion, distinct responsiveness ought to be considered throughout the operations of skidding to curtail unfriendly influences on soil physical properties in ground-base skidding.

Design, synthesis and photophysical studies of BODIPY-o, m, p-phenylenediamine-based probes: Insights into their responsiveness under different pH conditions

In this paper, a series of novel phenylenediamine-fluoroboron pyrrole fluorescent derivatives were prepared which have distinct responsiveness under different hydrogen ion concentration (pH) conditions. It is noticed that the products showed excellent fluorescence properties in different solvents, especially in tetrahydrofuran and dichloromethane, with the most prominent fluorescence intensity, while the fluorescence in methanol, acetonitrile, and dimethyl sulfoxide was weaker. Responsiveness under different hydrogen ion concentration conditions in aqueous solutions were also observed, where the fluorescence intensity is quenching when the pH is 4.0. With regard to cells imaging investigation, the products showed the prominent fluorescence in HeLa cells. Further acidic cell imaging results showed that under acidic conditions made of formic acid or acetic acid, the intracellular fluorescence of the compounds was clustered around the cells and intensive enough different from without acidity control group. Especially, the compounds have unique fluorescence in acidic environment and has great potential and research value as acidic probes.

Distinct Responsiveness of Tumor-Associated Macrophages to Immunotherapy of Tumors with Different Mechanisms of Major Histocompatibility Complex Class I Downregulation.

Simple SummaryTumor-associated macrophages (TAMs) are one of the major cell subpopulations in the tumor microenvironment (TME) where they can either be pro-tumorigenic or contribute to an anti-tumor immunity. The TME and TAM phenotype were analyzed after combined immuno-therapy (IT) in tumor models characterized by distinct expression of major histocompatibility class I complex (MHC-I) molecules, i.e., tumors induced with TC-1 (MHC-I-proficient), TC-1/A9 (reversibly downregulated), and TC-1/dB2m (irreversibly downregulated) cells. We found out that combined IT highly activated immune reactions in the TME of TC-1 and TC-1/A9 tumors, but the TME of TC-1/dB2m tumors remained almost unchanged. Correspondingly, TAMs from TC-1/A9 tumors were able to destroy tumor cells in vitro, while TAMs isolated from TC-1/dB2m tumors showed profoundly decreased cytotoxicity. Hence, various capabilities of TAMs in tumors with distinct expression of MHC-I molecules should be considered when applying IT, particularly IT focused on TAMs.Tumor-associated macrophages (TAMs) plentifully infiltrate the tumor microenvironment (TME), but their role in anti-tumor immunity is controversial. Depending on the acquired polarization, they can either support tumor growth or participate in the elimination of neoplastic cells. In this study, we analyzed the TME by RNA-seq and flow cytometry and examined TAMs after ex vivo activation. Tumors with normal and either reversibly or irreversibly decreased expression of major histocompatibility complex class I (MHC-I) molecules were induced with TC-1, TC-1/A9, and TC-1/dB2m cells, respectively. We found that combined immunotherapy (IT), composed of DNA immunization and the CpG oligodeoxynucleotide (ODN) ODN1826, evoked immune reactions in the TME of TC-1- and TC-1/A9-induced tumors, while the TME of TC-1/dB2m tumors was mostly immunologically unresponsive. TAMs infiltrated both tumor types with MHC-I downregulation, but only TAMs from TC-1/A9 tumors acquired the M1 phenotype upon IT and were cytotoxic in in vitro assay. The anti-tumor effect of combined IT was markedly enhanced by a blockade of the colony-stimulating factor-1 receptor (CSF-1R), but only against TC-1/A9 tumors. Overall, TAMs from tumors with irreversible MHC-I downregulation were resistant to the stimulation of cytotoxic activity. These data suggest the dissimilarity of TAMs from different tumor types, which should be considered when utilizing TAMs in cancer IT.

High BRAF variant allele frequencies are associated with distinct pathological features and responsiveness to target therapy in melanoma patients

BackgroundBRAF mutant melanoma patients are commonly treated with anti-BRAF therapeutic strategies. However, many factors, including the percentage of BRAF-mutated cells, may contribute to the great variability in patient outcomes.Patients and methodsThe BRAF variant allele frequency (VAF; defined as the percentage of mutated alleles) of primary and secondary melanoma lesions, obtained from 327 patients with different disease stages, was assessed by pyrosequencing. The BRAF mutation rate and VAF were then correlated with melanoma pathological features and patients’ clinical characteristics. Kaplan–Meier curves were used to study the correlations between BRAF VAF, overall survival (OS), and progression-free survival (PFS) in a subset of 62 patients treated by anti-BRAF/anti-MEK therapy after metastatic progression.ResultsA highly heterogeneous BRAF VAF was identified (3%-90%). Besides being correlated with age, a higher BRAF VAF level was related to moderate lymphocytic infiltration (P = 0.017), to melanoma thickness according to Clark levels, (level V versus III, P = 0.004; level V versus IV, P = 0.04), to lymph node metastases rather than cutaneous (P = 0.04) or visceral (P = 0.03) secondary lesions. In particular, a BRAF VAF >25% was significantly associated with a favorable outcome in patients treated with the combination of anti-BRAF/anti-MEK drug (OS P = 0.04; PFS P = 0.019), retaining a significant value as an independent factor for the OS and the PFS in the multivariate analysis (P = 0.014 and P = 0.003, respectively).ConclusionThese results definitively support the role of the BRAF VAF as a potential prognostic and predictive biomarker in melanoma patients in the context of BRAF inhibition.

Abstract PS18-41: Somatic mutational landscapes of invasive ductal and lobular carcinomas in the GENIE consortium cohort: Real-world gene actionability assessment of 8,756 breast cancer patients

Abstract Background: Invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) of the breast typically present distinct clinicopathological characteristics and responsiveness to systemic therapy. In addition, breast cancer data from The Cancer Genome Atlas (TCGA) have shown these two pathological subtypes also present distinct genomic features when analyzed using DNA copy number arrays and whole exome sequencing platforms. More recently, the AACR Project GENIE Consortium, which is a publicly accessible international cancer registry of real-world data assembled through data sharing among leading cancer centers in the world, have allowed in-depth analyses of clinical actionability using patient-level data from clinical next-generation sequencing (NGS) assays. In this study, we assessed the somatic mutational landscapes of a large cohort (n = 8,756) of invasive breast carcinomas from 19 institutions participating in the GENIE Consortium Cohort (v8.0) and examined clinical actionability of unique mutations identified in each breast cancer subtype. Method: We assessed the eighth data release of the GENIE Consortium Cohort encompassing targeted sequencing data from 7,647 IDC and 1,109 ILC cases. Clinical features and somatic mutations including single-nucleotide variants, small indels, fusions, and copy number alterations (CNAs) were retrieved from cBioportal and SAGE Bionetworks. Gene actionability was examined using both OncoKB and CiVIC publicly available knowledgebases. All patient samples were de-identified and encoded with GENIE sample codes. Results: Patients with IDC tumors were 5 years younger than patients with ILC tumors at the time sequencing data was reported (median 55 versus median 60 years old, Kruskal-Wallis, p < 10e-10). Both IDC and ILC had on average 2 mutations per tested sample. Overall, IDC and ILC tumors had median fractions of 22% and 14% of their genomes altered, respectively (Kruskal-Wallis, p < 10e-10). An initial gene enrichment analysis including 938 genes with point mutations and small indels identified CDH1 [Log-ratio (LR) 4.66, p < 1e-10], RHOA (LR 2.81, p = 1.3e-10), PTK2B (LR 2.68, p = 5.2e-4), ERBB2 (LR 1.80, p < 1e-10), TBX3 (LR 1.72, p < 1e-10), FOXA1 (LR 1.49, p = 2.5e-10) and RUNX1 (LR 1.25, p = 3.1e-9) as genes significantly enriched in ILC tumors. On the other hand, mutations in GATA3 (LR = 1.67, p < 1e-10) and TP53 (LR = 1.55, p < 1e-10) were significantly enriched in IDC tumors. A further gene enrichment analysis for copy-number alterations in 1139 genes showed amplification in PARP1 (LR 1.55 p = 2.5e-3) and deep deletions in IKZF1 (LR 2.8, p = 2.2e-3) and CDH1 (LR = 1.88, p = 1.7e-4) as the most enriched genes with CNAs in ILC. In parallel, amplifications in AURKA (LR 3.2, p = 1e-8), PPM1D (LR 3.1, p <1e-10), RAD51C (LR 2.85, p = 3.8e-8), BRIP1 (LR 2.66, p < 1e-10), ERBB2 (LR 1.65, p = 1e-10), MYC (LR 1.64, p = 1e-10), CDK12 (LR 1.55, p = 2.6e-9), and COL22A1 (LR 1.19, p = 1.6e-5), and deep deletion in CDKN2A (LR 2.1, p = 1.9e-6) were enriched in IDC tumors. Among those enriched alterations for each histological subtype, the knowledgebase CiVIC did not present curated data available for genes TBX3, FOXA1, GATA3, COL22A1, BRIP1, PPM1D, and RAD51C. OncoKB only missed genes PTK2B and COL22A1. Conclusions: Real-world genomic data from the GENIE Consortium Cohort support that breast cancer presents distinct mutational landscapes for IDC and ILC tumors. For each histological subtype, we confirmed there are different levels of enrichments for shared mutations in actionable genes. Despite the fact that publicly available knowledgebases present comprehensive curated information about commonly mutated genes in cancer, we noticed that actionability data of important cancer driver genes were missed. Citation Format: Alessandro Leal, Patricia Taranto, Poliana BG Blasi, Carlos Tadeu Garrote, Fernando Moura. Somatic mutational landscapes of invasive ductal and lobular carcinomas in the GENIE consortium cohort: Real-world gene actionability assessment of 8,756 breast cancer patients [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS18-41.

BRAF fusions in pediatric histiocytic neoplasms define distinct therapeutic responsiveness to RAF paradox breakers.

Pediatric histiocytic neoplasms are hematopoietic disorders frequently driven by the BRAF-V600E mutation. Here, we identified two BRAF gene fusions (novel MTAP-BRAF and MS4A6A-BRAF) in two aggressive histiocytic neoplasms. In contrast to previously described BRAF fusions, MTAP-BRAF and MS4A6A-BRAF do not respond to the paradox breaker RAF inhibitor (RAFi) PLX8394 due to stable fusion dimerization mediated by the N-terminal fusion partners. This highlights a significant and clinically relevant shift from the current dogma that BRAF-fusions respond similarly to BRAF-inhibitors. As an alternative, we show suppression of fusion-driven oncogenic growth with the pan-RAFi LY3009120 and MEK inhibition.

Fallopian tube epithelial cells express androgen receptor and have a distinct hormonal responsiveness when compared with endometrial epithelium.

How does steroid receptor expression, proliferative activity and hormone responsiveness of the fallopian tube (FT) epithelium compare to that of the endometrial epithelium? Proliferative indices, hormone receptor expression-scores and in vitro response to oestrogen and androgens of the human FT demonstrate a distinct pattern from the matched endometrium. The FT epithelium exists as a continuum of the endometrium, and both express steroid hormone receptors. The ovarian steroid hormones regulate cyclical proliferation and regeneration of the endometrium, but their effects on steroid hormone receptor expression and proliferation in the FT have not yet been fully elucidated. We included women with proven fertility, undergoing hysterectomy and bilateral salpingo-oophorectomy for benign, gynaecological conditions at Liverpool Women's NHS Foundation Trust. They had no known endometrial or tubal pathology and were not on hormonal treatments for at least 3 months preceding sample collection in this prospective observational study (conducted between 2010 and 2018). A full-thickness sample of the endometrium and a sample from the FT were collected from each woman. The differential protein and mRNA levels of steroid hormone receptors, oestrogen receptors α and β, androgen receptor (AR) and progesterone receptor (PR), and the proliferative marker (Ki67) of the endometrium and the FT tissue samples from 47 healthy women undergoing surgery (37 premenopausal and 10 postmenopausal) were investigated using immunohistochemistry and quantitative real-time PCR. The comparative responsiveness to oestrogen and androgen of the endometrium and the fimbrial end of the FT was analysed using an in vitro short-term explant culture model. The endpoints assessed in the explants were the changes in mRNA and protein levels for AR, PR and the epithelial proliferative index after 24 h treatment with oestradiol (E2) or dihydrotestosterone (DHT). The premenopausal endometrial functionalis glands (FG) displayed the well-known cyclic variation in cellular proliferation and steroid receptor scores. Compared with the endometrial FG, the matched FT epithelium (both fimbrial or isthmic ends) displayed a significantly lower proportion of cells expressing Ki67 (2.8% ± 2.2%, n = 18 vs 30.0% ± 26.3%, n = 16, P = 0.0018, respectively) accompanied with a significantly higher AR immunoscores (6.7 ± 2.7, n = 16 vs 0.3 ± 1.0, n = 10, P = 0.0136). The proportion of cells expressing Ki67 and the AR immunoscores of the FT epithelium correlated positively with endometrial luminal epithelium (r = 0.62, P = 0.005, and r = 0.68, P = 0.003, respectively). In vitro experiments suggested the tubal explants to be apparently less responsive to E2 yet more sensitive to DHT compared with the matched endometrium explants. The short-term in vitro nature of the tissue explant cultures used in the study may not be representative of how different anatomical regions of the endometrium and FT behave in vivo. Our study included a high proportion of older premenopausal women with a regular menstrual cycle, which may therefore affect extrapolation of findings to a younger group. Advancing our understanding of tubal and endometrial epithelial cell function has important implications for the diagnosis and treatment of diseases such as infertility, ectopic pregnancy, endometriosis and cancer. The work included in this article was funded by Wellbeing of Women project grants RG1073 and RG2137 (D.K.H.) and Wellbeing of Women Entry-Level Scholarship ELS706 (A.M). A.M. was also supported by an NIHR ACF fellowship grant. Further support received from Liverpool Women's Hospital NHS Trust (S.M.), University of Liverpool (E.B. and A.W.). All authors declare there are no conflicts of interest. N/A.

Abstract 632: Novel BRAF gene fusions in pediatric histiocytic neoplasms respond differentially to RAF targeted therapies based on dimerization profiles

Abstract Introduction: Pediatric histiocytic neoplasms are clonal hematopoietic disorders driven by mutations activating the mitogen-activated protein kinase (MAPK) pathway, such as BRAF- V600E. In non-BRAFV600E cases, we investigated alternative MAPK mutations and found two novel BRAF gene fusions. We have previously shown effective targeting of BRAF fusions (KIAA1549-BRAF) in pediatric low-grade gliomas (PLGGs) with paradox breaker RAF inhibitor (RAFi), PLX8394 and MEK inhibitors (MEKi). Here, we investigate the distinct responsiveness of novel BRAF fusions to RAFi therapies and explore the mechanistic basis of such differential responses compared to other BRAF fusions. Method: Two histiocytic patient tumors were analyzed using the CHOP Comprehensive Next- Gen Sequencing Solid Tumor Panel and a targeted RNA-seq panel for 106 fusion partner genes. Novel BRAF fusions identified were sub-cloned to create heterologous cell models. Immunoblotting of serum starved cells measured MAPK and PI3K pathway activation and soft agar assay tested for oncogenic phenotypes driven by BRAF fusions, along with response to PLX8394, LY3009120 (pan-RAF dimer inhibitor), and MEKi. Co-immunoprecipitation assays using Myc- and Flag-tagged BRAF fusion constructs assessed dimerization profiles, with or without inhibitors. Result: In the two M- and L-type histiocytic neoplasms assessed, we found novel and rare BRAF gene fusions, MTAP-BRAF and MS4A6A-BRAF, respectively. Both BRAF fusions activated the MAPK/ PI3K pathways and showed homo- and hetero-dimerization with BRAF and the respective N-terminal fusion partner. Compared to common BRAF fusions, MTAP-BRAF and MS4A6A- BRAF did not respond to PLX8394 due to no disruption of active fusion homo- and hetero-dimers, which was in turn was due to the untargeted, stable dimerization mediated by the N-terminal fusion partners. Conversely, we observed robust suppression with LY3009120 that bound fusion dimers and kept them in an inactivate confirmation. MEKi selumetinib and trametinib also suppressed fusion driven signaling and oncogenic phenotypes. Conclusion: We show that novel MTAP-BRAF and MS4A6A-BRAF fusions, found in histiocytic tumors, do not respond to RAFi PLX8394 but are targeted by LY3009120 or MEKi. Our finding that PLX8394 does not disrupt MTAP-BRAF or MS4A6A-BRAF dimerization due to contribution of N-terminal partners defines a novel paradigm for the distinct mechanisms sought by BRAF fusions in response to RAFi therapy. We show potent suppression of these mechanistically distinct BRAF fusions with MEKi or LY3009120 which function independent of vulnerability to fusion dimerization. Overall, this study highlights the unique and differential biology hijacked by BRAF fusions in response to RAFi and further warrants detailed mechanistic classification of BRAF fusions based on their responsiveness to targeted agents. Citation Format: Payal Jain, Lea F. Surrey, Joshua Straka, Pierre Russo, Richard Womer, Marilyn Li, Phillip B. Storm, Angela J. Waanders, Michael Hogarty, Jennifer Picarsic, Adam C. Resnick. Novel BRAF gene fusions in pediatric histiocytic neoplasms respond differentially to RAF targeted therapies based on dimerization profiles [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 632.

Root suckering promotes recruitment in two temperate rainforest trees with contrasting shade tolerance

The regeneration niche differentiation helps to explain plant coexistence and thus biodiversity. The study of the regeneration niche has been traditionally based on sexual recruitment, while overlooking clonal growth. Root suckering offers a successful alternative for local dispersal under suboptimal conditions for sexual reproduction. For light-limited forests, we hypothesized that: 1) root suckering would increase the regeneration niche towards high-light conditions in shade-tolerant trees and towards dark conditions in light-demanding species; 2) contrasting responses of survival and growth to light availability would explain niche differentiation of both suckers and saplings; and 3) distinct responsiveness to light among species and recruit-types would reflect differences in functional traits. We tested these hypotheses with two evergreen tree species that coexist in the temperate rainforest of southern South America: Embothrium coccineum (light-demanding) and Eucryphia cordifolia (shade-tolerant). We measured the light availability in two study plots above each recruit and along transects established in the understory. Niche selection, niche differentiation and changes in survival probability with light were inferred from the analysis of the light frequency distributions. We evaluated the effect of light on the relative volumetric change in stems over a 1-year period. Functional traits of leaves, stems, and crowns were measured in suckers and saplings growing under similar light conditions; these traits were then compared among size classes, recruit-types and species. Root suckering was the prevalent reproduction mode of both studied species, extending the light niche towards open microenvironments only during the earliest ontogenetic stages. The poor structural strength of the leaves and wood of small Eucryphia saplings explains its underuse of open microsites. Neither photosynthetic assimilation nor carbon subsidy can sustain Embothrium suckers at the shadiest microsites. Suckering proved to increase the persistence of Embothrium until advanced stages of forest succession, facilitating its coexistence with the late-successional Eucryphia. Our study emphasizes that clonal growth is essential to understand the dynamics of temperate rainforests.

Stretchable Sensors with Tunability and Single Stimuli-Responsiveness through Resistivity Switching Under Compressive Stress.

The fascinating human somatosensory system with its complex structure is composed of numerous sensory receptors possessing distinct responsiveness to stimuli. It is a continuous source of inspiration for tactile sensors that mimic its functions. However, to achieve single stimulus-responsiveness with mechanical decoupling is particularly challenging in the light of structural design and has not been fully addressed to date. Here we propose a novel structural design inspired by combining the characteristics of electronic skin (e-skin) and electronic textile (e-textile) into a hybrid interface to achieve a stretchable single stimuli-responsive tactile sensor. The stencil printable biocarbon composite/silver-plated nylon hybrid interface possesses an extraordinary resistance switching (ΔR/R0 up to ∼104) under compressive stress which is controllable by the composite film-thickness. It achieves a very high normal pressure sensitivity (up to 60.8 kPa–1) in a wide dynamic range (up to ∼50 kPa) in the piezoresistive operation mode and can effectively decouple stresses induced by stretching or bending. In addition, the device is capable of high accuracy strain sensing in its capacitive operation mode through dimensional change dominant response. Because of these intriguing features, it has potential for the next-generation Internet of Things devices and user-interactive systems capable of providing visual feedback and more advanced robotics or even prosthetics.

Phenanthroline-Based Polyarylate Porous Membranes with Rapid Water Transport for Metal Cation Separation.

The selective separation of ions in terms of extremely similar size and properties remains an important challenge in water purification. We innovated a kind of porous nanofilm via interfacial polymerization using rigid heterocyclic ligands to achieve high valent cation selectivity and rapid water/ion transport. The interconnected microporosity and uniformly distributed cation-affinitive sites of the ultrathin membranes enabled water permeation (7.5 L m-2 h-1 bar-1), ion permeance of Na+ (1.5 mol m-2 h-1 bar-1), and Mg2+/Na+ permselectivity (2.1) during nanofiltration. The forward osmosis exhibited a prominent water flux of 95 LMH at 1 M NaCl draw solution, which expanded various applications. The polyarylate membranes comprising 4,7-diphenyl-1,10-phenanthroline showed a higher water permeation and ion selectivity than the planar monomers, e.g., resorcinol. A distinct fluorescence responsiveness existed between membranes and cations for the interaction characterization. Host-guest nuclear magnetic resonance (NMR) spectroscopy and solid-state nuclear magnetic resonance spectroscopy characterized the preferential affinitive of divalent/high-valent cations in the interconnected microporous powders; an ultraviolet spectrophotometer characterized the light responsiveness of the porous nanofilms. Such an active membrane has potential applications in selective separation and adsorption of cations, photocatalytic materials, photosensors, and other fields.