Histological Phenotype Research Articles

First and second morning spot urine protein measurements for the assessment of proteinuria: a diagnostic accuracy study in kidney transplant recipients

BackgroundQuantification of proteinuria in kidney transplant recipients is important for diagnostic and prognostic purposes. Apart from correlation tests, there have been few evaluations of spot urine protein measurements in kidney transplantation.MethodsIn this cross-sectional study involving 151 transplanted patients, we investigated measures of agreement (bias and accuracy) between the estimated protein excretion rate (ePER), determined from the protein-to-creatinine ratio in the first and second morning urine, and 24-h proteinuria and studied their performance at different levels of proteinuria. Measures of agreement were reanalyzed in relation to allograft histology in 76 patients with kidney biopsies performed for cause before enrolment in the study.ResultsFor ePER in the first morning urine, percent bias ranged from 1 to 28% and accuracy (within 30% of 24-h collection) ranged from 56 to 73%. For the second morning urine, percent bias ranged from 2 to 11%, and accuracy ranged from 71 to 78%. The accuracy of ePER (within 30%) in first and second morning urine progressively increased from 56 and 71% for low-grade proteinuria (150–299 mg/day) to 60 and 74% for moderate proteinuria (300–999 mg/day), and to 73 and 78% for high-grade proteinuria (≥1000 mg/day). Measures of agreement were similar across histologic phenotypes of allograft injury.ConclusionsThe ability of ePER to accurately predict 24-h proteinuria in kidney transplant recipients is modest. However, accuracy improves with an increase in proteinuria. Given the similar accuracy of ePER measurements in first and second morning urine, second morning urine can be used to monitor protein excretion.

Immune-related histologic phenotype in pretreatment tumor biopsy predicts pathologic response to neoadjuvant anti-PD-1 treatment in squamous lung cancer.

e20540 Background: The neoadjuvant platform affords a rich and valuable resource for understanding the responses to therapy and carrying out reverse translation, including pathologic morphology. We aimed to develop a pretreatment histologic scoring system reflecting the preexisting immune response to predict the efficacy of neoadjuvant immunotherapy based on the morphological changes we mastered in the pathologic assessment after neoadjuvant immunotherapy. Methods: Surgical specimens from the 31 squamous cell lung cancer patients recruited in a phase Ib study of neoadjuvant anti-PD-1 therapy and eligible paired pretreatment biopsies from 15 of them were included in this study. The posttreatment surgical specimens were assessed according to the immune-related pathologic response criteria. Immune-related histologic phenotype assessment criteria (irHPC) were developed based on the pathologic features identified after neoadjuvant anti-PD-1 treatment. Three pathologists trained for irHPC independently scored the HE slides of the 15 pretreatment tumor biopsies according to irHPC. Results: Whether necrosis was included in the calculation of percent of residual viable tumor (%RVT) or not had almost no effect on the consistency of pathologic assessment ( P= 0.811) and the histological response grouping. The inter-pathologist variability of assessing %RVT with immune-activated phenotype was not statistically significant ( P= 0.480). Four immune-related features of pretreatment biopsies were included for calculating the predictive score, including three positive features (tumor-infiltrating lymphocytes, tumor-infiltrating eosinophils and dense plasma cells in stroma) and one negative feature (tumor-infiltrating neutrophils) according to the developed irHPC scoring system. The trained pathologist accurately predicted 6 out of 8 patients in the cPR/MPR group and 5 out of 7 patients in the non-cPR/MPR group according to irHPC. For inter-observer reproducibility using “2 points” as the cut-off point, the overall percent agreement (OPA) was 77.8%. The reliability between pathologists for a binary tumor evaluation showed “moderate” agreement (κ = 0.54). Conclusions: The irHPC scoring system reflecting the preexisting immune response could be used to predict the pathologic response of neoadjuvant immunotherapy, but still needs the larger trails to verify.

Differentiated activities of decorin and biglycan in the progression of post-traumatic osteoarthritis

To delineate the activities of decorin and biglycan in the progression of post-traumatic osteoarthritis (PTOA). Three-month-old inducible biglycan (BgniKO) and decorin/biglycan compound (Dcn/BgniKO) knockout mice were subjected to the destabilization of the medial meniscus (DMM) surgery to induce PTOA. The OA phenotype was evaluated by assessing joint structure and sulfated glycosaminoglycan (sGAG) staining via histology, surface collagen fibril nanostructure and calcium content via scanning electron microscopy, tissue modulus via atomic force microscopy-nanoindentation, as well as subchondral bone structure and meniscus ossification via micro-computed tomography. Outcomes were compared with previous findings in the inducible decorin (DcniKO) knockout mice. In the DMM model, BgniKO mice developed similar degree of OA as the control (0.44 [-0.18 1.05] difference in modified Mankin score), different from the more severe OA phenotype observed in DcniKO mice (1.38 [0.91 1.85] difference). Dcn/BgniKO mice exhibited similar histological OA phenotype as DcniKO mice (1.51 [0.97 2.04] difference vs control), including aggravated loss of sGAGs, salient surface fibrillation and formation of osteophyte. Meanwhile, Dcn/BgniKO mice showed further cartilage thinning than DcniKO mice, resulting in the exposure of underlying calcified tissues and aberrantly high surface modulus. BgniKO and Dcn/BgniKO mice developed altered subchondral trabecular bone structure in both Sham and DMM groups, while DcniKO and control mice did not. In PTOA, decorin plays a more crucial role than biglycan in regulating cartilage degeneration, while biglycan is more important in regulating subchondral bone structure. The two have distinct activities and modest synergy in the pathogenesis of PTOA.

Radiomics of Gadoxetate Disodium-Enhanced MRI in Glypican 3-Positive Hepatocellular Carcinoma Identification and Prognosis Evaluation

Background: To investigate the impact of preoperative gadoxetate disodium (Gd-EOB-DTPA) MRI-based radiomics on predicting glypican 3 (GPC3) and recurrence-free survival (RFS) of hepatocellular carcinoma (HCC). Methods: Between January 2014 and October 2018, 259 patients with solitary HCC ≤5 cm who underwent liver resection and preoperative Gd-EOB-DTPA MRI were recruited. Draw support from 5-fold cross-validation, the best of 15 radiomics models obtained by combining five feature selection strategies and three classifiers was integrated with independent clinicoradiologic predictors into the comprehensive nomogram. Findings: GPC3 was an independent risk factor for postoperative recrudescence in patients with solitary HCC below 5 cm. Alpha-fetoprotein >20 ng/mL, homogenous T2-weighted imaging and hypointensity on hepatobiliary phase were independently vulnerable to GPC3 in the clinicoradiologic model. With ten features selected by support vector machines-recursive feature elimination, logistic regression-based classifier achieved the best performance among 15 radiomics models. After 5-fold cross-validation, our comprehensive nomogram acquired better average area under receiver operating characteristic curves with 95% confidence intervals (training and validation cohorts: 0.931[0.889-0.972] Vs. 0.943[0.880-0.998]) than the clinicoradiologic algorithm (0.738[0.651-0.824] Vs. 0.739[0.563-0.915]) and the optimal radiomics model (0.943[0.910-0.978] Vs. 0.931[0.860-0.989]). Net reclassification indexes further demonstrated the superiority of GPC3 nomogram over the clinicoradiologic and radiomics algorithms(46.54%, P 77.9 to 48.2 months, P=0.044), which was analogue to that of the histological GPC3-positive phenotype (from >73.9 to 43.2 months, P<0.001). Interpretation: Preoperative radiomics-based nomogram of Gd-EOB-DTPA MRI satisfactorily distinguished GPC3 status and outcomes of patients suffered from solitary HCC within 5 cm. Funding: This research was supported by National Natural Science Foundation of China (No.91859107), Zhongshan Hospital, Fudan University (No. 2018ZSLC22), Shanghai Municipal Key Clinical Specialty (No.W2019-018), and Clinical Research Plan of SHDC (No.SHDC2020CR1029B). The funding agency played no role in this study. Declaration of Interest: The authors declare no potential conflict of interest. Ethical Approval: This retrospective study was approved and granted an exemption from written informed consent by the ethics committee of our hospital (No. B2021-113).

Combined Beneficial Effect of Voluntary Physical Exercise and Vitamin D Supplementation in Diet-induced Obese C57BL/6J Mice.

Physical exercise (PE) combined with nutritional approaches has beneficial effects that are widely advocated to improve metabolic health. Here we used voluntary PE together with vitamin D (VD) supplementation, which has already shown beneficial effects in primary and tertiary prevention in obese mice models, to study their combined additive effects on body weight management, glucose homeostasis, metabolic inflammation, and liver steatosis as key markers of metabolic health. Ten-week-old male C57BL/6J mice were fed a high-fat/sucrose (HFS) diet for 10 wk, then assigned to a 15-wk intervention period with PE, VD supplementation, or both PE and VD supplementation. Morphological, histological, and molecular phenotype data were characterized. The HFS-induced increases in body mass, adiposity, and adipocyte hypertrophy were improved by PE but not by VD supplementation. The HFS-induced inflammation (highlighted by chemokines mRNA levels) in inguinal adipose tissue was decreased by PE and/or VD supplementation. Furthermore, the intervention combining PE and VD showed additive effects on restoring insulin sensitivity and improving hepatic steatosis, as demonstrated through a normalization of size and number of hepatic lipid droplets and triglyceride content and a significant molecular-level decrease in the expression of genes coding for key enzymes in hepatic de novo lipogenesis. Taken together, our data show beneficial effects of combining PE and VD supplementation on obesity-associated comorbidities such as insulin resistance and hepatic disease in mice. This combined exercise-nutritional support strategy could prove valuable in obesity management programs.

Genome-wide association studies demonstrate that TASP1 contributes to increased muscle fiber diameter.

Muscle fiber diameter is an economically important trait because it affects meat yield and quality. However, the genetic basis underlying muscle fiber diameter has not been determined. In this study, we collected THREE muscular histological phenotypes in 479 ducks from an F2 segregating population generated by mallard × Pekin duck crosses. We performed genome-wide association studies (GWAS) and identified a quantitative trait locus (QTL) significantly associated with muscle fiber diameter on chromosome 3. Then, we discovered the selection signatures using the fixation index among 40 mallards and 30 Pekin ducks in this QTL region. Furthermore, we characterized the recombination event in this QTL region and identified a 6-kb block located on TASP1 that was significantly associated with muscle fiber diameter. Finally, five SNPs were screened as potential causative mutations within the 6-kb block. In conclusion, we demonstrated that TASP1 contributes to an increase in muscle fiber diameter, which helps to characterize muscle development and contributes to the genetic improvement of meat yield and quality in livestock.

Association of non-HLA antibodies against endothelial targets and donor-specific HLA antibodies with antibody-mediated rejection and graft function in pediatric kidney transplant recipients

BackgroundNon-HLA antibodies against endothelial targets have been implicated in the pathogenesis of antibody-mediated rejection (ABMR), but data in pediatric patients are scarce.MethodsWe retrospectively analyzed a carefully phenotyped single-center (University Children’s Hospital Heidelberg, Germany) cohort of 62 pediatric kidney transplant recipients (mean age at transplantation, 8.6 ± 5.0 years) at increased risk of graft function deterioration. Patients had received their transplant between January 1, 1999, and January 31, 2010. We examined at time of late index biopsies (more than 1-year post-transplant, occurring after January 2004) the association of antibodies against the angiotensin II type 1 receptor (AT1R), the endothelin type A receptor (ETAR), the MHC class I chain-like gene A (MICA), and vimentin in conjunction with overall and complement-binding donor-specific HLA antibodies (HLA-DSA) with graft histology and function.ResultsWe observed a high prevalence (62.9%) of non-HLA antibody positivity. Seventy-two percent of HLA-DSA positive patients showed additional positivity for at least one non-HLA antibody. Antibodies against AT1R, ETAR, and MICA were associated with the histological phenotype of ABMR. The cumulative load of HLA-DSA and non-HLA antibodies in circulation was related to the degree of microinflammation in peritubular capillaries. Non-HLA antibody positivity was an independent non-invasive risk factor for graft function deterioration (adjusted hazard ratio 6.38, 95% CI, 2.11–19.3).ConclusionsOur data indicate that the combined detection of antibodies to HLA and non-HLA targets may allow a more comprehensive assessment of the patients’ immune responses against the kidney allograft and facilitates immunological risk stratification.

Altered Visual Function in a Larval Zebrafish Knockout of Neurodevelopmental Risk Gene pdzk1.

PurposeThe human PDZK1 gene is located in a genomic susceptibility region for neurodevelopmental disorders. A genome-wide association study identified links between PDZK1 polymorphisms and altered visual contrast sensitivity, an endophenotype for schizophrenia and autism spectrum disorder. The PDZK1 protein is implicated in neurological functioning, interacting with synaptic molecules including postsynaptic density 95 (PSD-95), N-methyl-d-aspartate receptors (NMDARs), corticotropin-releasing factor receptor 1 (CRFR1), and serotonin 2A receptors. The purpose of the present study was to elucidate the role of PDZK1.MethodsWe generated pdzk1-knockout (pdzk1-KO) zebrafish using CRISPR/Cas-9 genome editing. Visual function of 7-day-old fish was assessed at behavioral and functional levels using the optomotor response and scotopic electroretinogram (ERG). We also quantified retinal morphology and densities of PSD-95, NMDAR1, CRFR1, and serotonin in the synaptic inner plexiform layer at 7 days, 4 weeks, and 8 weeks of age. Standard RT-PCR and nonsense-mediated decay interference treatment were also performed to assess genetic compensation in mutants.ResultsRelative to wild-type, pdzk1-KO larvae showed spatial frequency tuning functions with increased amplitude (likely due to abnormal gain control) and reduced ERG b-waves (suggestive of inner retinal dysfunction). No synaptic phenotypes, but possible morphological retinal phenotypes, were identified. We confirmed that the absence of major histological phenotypes was not attributable to genetic compensatory mechanisms.ConclusionsOur findings point to a role for pdzk1 in zebrafish visual function, and our model system provides a platform for investigating other genes associated with abnormal visual behavior.

Histologic Phenotypes and Outcomes in Single vs Double Lung Transplantation among Recipients with Interstitial Lung Disease

<h3>Purpose</h3> The role of single vs double lung transplantation for ILD remains controversial due to heterogeneity within this population. This study elucidates the role of explant histology on outcomes among patients undergoing a single vs double lung transplant. <h3>Methods</h3> A total of 412 patients with ILD transplanted from 2001-2019 at our institution were retrospectively reviewed. Patients were stratified by those who underwent a single (n=239, 58%) or double (n=173, 42%) lung transplant. Explant pathology reports were analyzed to understand the effect of histologic phenotypes and post-transplant outcomes. Primary outcomes of interest included overall and CLAD free survival. Treatment effects were obtained using a multivariable Cox-proportional hazards function. <h3>Results</h3> Of the total population, 41.7% had UIP, 26.5% NSIP, 5.8% mixed NSIP+UIP, 14.8% hypersensitivity pneumonitis (HP), and 11.2% sarcoid on explant pathology. Overall survival was similar between histologic phenotypes. HP and NSIP had earlier development of CLAD compared to sarcoidosis and UIP. A single lung transplant was an independent predictor of mortality among recipients with HP at explant pathology on multivariable analysis. Single lung transplant was also an independent predictor for CLAD among patients with NSIP and HP. The rest of the histologic phenotypes UIP, mixed NSIP+UIP ad sarcoid failed to show a significant difference in any of the outcomes by the use of single vs double lung transplant. <h3>Conclusion</h3> Patients with ILD have distinct pathologic characteristics and related post lung transplant outcomes. Recipients with UIP, mixed NSIP+UIP and sarcoid treated with a single lung transplant had no adverse overall and CLAD-free survival. Recipients with conditions at risk for earlier CLAD, such as HP and NSIP, may benefit from double lung transplantation. Lung biopsies prior to transplantation may aid in donor lung allocation among patients with ILD and optimize organ utilization.

Celiac disease antibody levels reflect duodenal mucosal damage but not clinical symptoms

Objectives This study aimed to investigate, in a real-world population, whether the histological and clinical phenotype differ at baseline and during follow-up in patients with high and low CD (celiac disease) antibody titers. Materials and methods The study cohort consisted of 96 consecutive patients diagnosed to have CD during the years 2010–2018. The clinical parameters, symptoms and laboratory results were registered and histomorphometry was analyzed from the available duodenal biopsies taken during the primary and follow-up esophageal-gastricduodenoscopies. Patients having immunoglobulin A transglutaminase antibody (tTG-ab) levels above 70 U/mL were classified as high titer patients. Results Measured by the villous-crypt ratio, the duodenal mucosa was more severely damaged in the high tTG-ab group than in the low tTG-group at baseline (n = 70, 0.61 ± 0.63 vs. 1.02 ± 0.87, p = .003) and during the follow-up when the patients were on gluten-free diet (n = 27, 1.80 ± 0.72 vs. 2.35 ± 0.64, p = .041). Interestingly, the high tTG-ab group members had fewer gastrointestinal symptoms at baseline than those in the low tTG-ab group (43% vs. 68%, p = .013) but lower vitamin D levels (68 ± 34 nmol/L vs. 88 ± 29 nmol/L, p = .034) and more often microcytosis (28% vs. 10%, p = .040). During the follow-up, these differences were no longer detected. Conclusions At baseline, CD patients with high tTG-ab have more severe duodenum injury and signs of malabsorption but fewer symptoms. After gluten-free diet has been initiated, the mucosal healing in the high tTG-ab group is prolonged, but symptoms and signs of malabsorption recover equally in both groups.

The potential role of plasma fibroblast growth factor 21 as a diagnostic biomarker for abdominal aortic aneurysm presence and development

AimsFibroblast growth factor 21 (FGF21) has been identified as the master hormonal regulator of energy balance, its elevation is observed in a series of metabolic and cardiovascular diseases. Studies have implicated the role of FGF21 signaling in the pathogenesis of abdominal aortic aneurysm (AAA). We will investigate the association of FGF21 and AAA development. Materials and methodsIn this study, we assayed plasma levels of FGF21 in 82 patients with AAA and 44 control subjects, then analyzed their relationship with clinical, biochemical and histological phenotypes. The expression of β-klotho, an essential co-receptor of FGF21, was assessed with IHC staining and RT-qPCR. Machine learning models incorporate a combination of FGF21 and clinical data were utilized in the prediction of AAA occurrence. Key findingsFGF21 was statistically higher in patients with AAA (781 pg/ml [533, 1213]) than in control subjects (567 pg/ml [324, 939]). After adjustment for age and BMI, we found a positive association of FGF21 levels with AAA diameters, hypertension rate and hsCRP, and a negative correlation between FGF21 levels and HDL-c. Furthermore, the protein levels of β-klotho in abdominal aorta of AAA were found significantly lower than in control group indicating the presence of FGF21 resistance. Combining FGF21 levels with four clinical characteristics significantly improved the stratification of AAA and control groups with an AUC of 0.778. SignificanceCombining detection of plasma FGF21 and clinical characteristics may be reliable for identifying the presence of AAA. The role of FGF21 as a therapeutic target of AAA warrants further investigation.

Deep learning classification of lung cancer histology using CT images

Tumor histology is an important predictor of therapeutic response and outcomes in lung cancer. Tissue sampling for pathologist review is the most reliable method for histology classification, however, recent advances in deep learning for medical image analysis allude to the utility of radiologic data in further describing disease characteristics and for risk stratification. In this study, we propose a radiomics approach to predicting non-small cell lung cancer (NSCLC) tumor histology from non-invasive standard-of-care computed tomography (CT) data. We trained and validated convolutional neural networks (CNNs) on a dataset comprising 311 early-stage NSCLC patients receiving surgical treatment at Massachusetts General Hospital (MGH), with a focus on the two most common histological types: adenocarcinoma (ADC) and Squamous Cell Carcinoma (SCC). The CNNs were able to predict tumor histology with an AUC of 0.71(p = 0.018). We also found that using machine learning classifiers such as k-nearest neighbors (kNN) and support vector machine (SVM) on CNN-derived quantitative radiomics features yielded comparable discriminative performance, with AUC of up to 0.71 (p = 0.017). Our best performing CNN functioned as a robust probabilistic classifier in heterogeneous test sets, with qualitatively interpretable visual explanations to its predictions. Deep learning based radiomics can identify histological phenotypes in lung cancer. It has the potential to augment existing approaches and serve as a corrective aid for diagnosticians.

Height in adolescence as a risk factor for glioma subtypes: a nationwide retrospective cohort study of 2.2 million subjects.

Gliomas manifest in a variety of histological phenotypes with varying aggressiveness. The etiology of glioma remains largely unknown. Taller stature in adulthood has been linked with glioma risk. The aim of this study was to discern whether this association can be detected in adolescence. The cohort included 2 223 168 adolescents between the ages of 16 and 19 years. Anthropometric measurements were collected at baseline. Incident cases of glioma were extracted from the Israel National Cancer Registry over a follow-up period spanning 47 635 745 person-years. Cox proportional hazard models were used to estimate the hazard ratio (HR) for glioma and glioma subtypes according to height, body mass index (BMI), and sex. A total of 1195 patients were diagnosed with glioma during the study period. Mean (SD) age at diagnosis was 38.1 (11.7) years. Taller adolescent height (per 10-cm increase) was positively associated with the risk for glioma of any type (HR: 1.15; P = .002). The association was retained in subgroup analyses for low-grade glioma (HR: 1.17; P = .031), high-grade glioma (HR: 1.15; P = .025), oligodendroglioma (HR: 1.31; P = .015), astrocytoma (HR: 1.12; P = .049), and a category of presumed IDH-mutated glioma (HR: 1.17; P = .013). There was a trend toward a positive association between height and glioblastoma, however this had borderline statistical significance (HR: 1.15; P = .07). After stratification of the cohort by sex, height remained a risk factor for men but not for women. The previously established association between taller stature in adulthood and glioma risk can be traced back to adolescence. The magnitude of association differs by glioma subtype.

Abstract PS16-12: Double heterozygous pathogenic variants prevalence in a cohort of hereditary breast cancer patients

Abstract Significant progress has been made in the identification of inherited genetic factors underlying hereditary cancers. Inherited pathogenic variants in cancer predisposition genes that confer moderate to high risk of breast cancer (BC) are found in 10% of BC cases. Pathogenic variants in BRCA1 and BRCA2 genes are the underlying alterations in hereditary BC. Women with a pathogenic variant in BRCA1 or BRCA2 are at substantially higher risk of developing breast, ovarian and pancreatic cancers and represent 25-28% of hereditary BC cases. The gold standard for molecular diagnosis of a hereditary cancer syndrome is aNext Generation Sequencing (NGS) panel that includes all genes known to confer moderate to high-risk for BC (TP53, PTEN, CHEK2, ATM, PALB2 STK11, RAD51C and BRIP1). Identifying who is eligible to perform genetic testing remains a challenge but presently age at the time of tumor diagnosis, family history and disease stage are still the best predictors. The identification of a germline pathogenic variant enables appropriate genetic counseling, risk-reducing interventions. The detection of variants in BRCA1, BRCA2 and PALB2 may also determine treatment options in patients with advanced or metastatic disease. The known non-BRCA1/2-associated hereditary BC comprise a heterogeneous group of tumors, for most of which the prevalence of histologic and immunohistochemical (IHQ) phenotypes are yet to be identified while triple-negative BC is the most prevalent in BRCA1 carriers. The co-occurence of more than one germline pathogenic variant in BC genes is a rare event and it may affect cancer risk and penetrance is still unknown. Only a few reports on cases and series of double mutation carriers have been described in BRCA1, BRCA2, PALB2, CHEK2, BLM or NBN. In this series case we report patients with double heterozygous (DH) mutation in BC, their age at diagnosis, presence of second neoplasia and cancer phenotype in moderate and high penetrance genes on germline panels. Considering this scenario, we performed an active search among the 2651 women from the Hereditary Cancer Registry at Hospital Sirio-Libanes, São Paulo, Brazil, between January 2013 and June 2020. We have included 229 patients with a histopathological confirmation of a BC who have tested positive for pathogenic/likely pathogenic variant. Of these, 4,3% (10/229) women were identified with DH mutation in high and moderate penetrance genes in BC (table 1). All patients carried a germline pathogenic variant in at least one high-penetrance gene. A total of 7 cases (7/10) carried at least one BRCA1/2 pathogenic variant. Two women (2/10) included had developed bilateral BC. The occurrence of a second primary cancer was described in three patients (3/10). The median age at first BC diagnosis was 36,7 years old. Two BC were found to be triple negative, five cases were of Luminal subtype and one was HER2+. The age at diagnosis is similar to most hereditary BC related to those developed by BRCA1 and TP53 germline variant carriers. In this case series there is no evidence of additive effect on the risk of tumor development. Larger series of DH mutation carriers are needed to estimate the risk in double heterozygous populations and to determine whether the presence of DH have a synergistic interaction or additive effect on cancer risk. Table1 - Breast Cancer double variantsBreast Cancer double VariantsGeneVariantAge of Breast Cancer onset IHQ statusBilateral BCOther prymary neoplasia (age)BRCA1c.5266dupC36 yoER- PR- HER2 +YesPancreas (40), Lung (53)TP53c.1010G&amp;gt;ABRCA1c.4165.4166delAG32yoN/ANoBreast (37), Skin (50)PALB2c.1240C&amp;gt;TBRCA1c.5266dupC33yoER - RP- HER2 -NoNoATMc.6729_6730delAABRCA1c.3748G&amp;gt;T33yoER- PR -HER2-NoNoRECQL4c.1568_1573delinsCCCCCBRCA2c.2516dupA49yoER+ PR+ HER2 -NoNoPMS2c.2182_2184delinsGBRCA2c.8960del32yoER+ PR+ HER2-NoNoATMc.901+1G&amp;gt;ABRCA1c.1687C&amp;gt;T38yoER+NoBreast Cancer (58)NBNc.1142delC PR +MUTYHc.536A&amp;gt;G Her2-TP53c.1010G&amp;gt;A35yoN/ANoNoMUTYHc.1187G&amp;gt;AATMc.4906C&amp;gt;T42yoER+NoNoTP53c.1010G&amp;gt;APR+HER2-PALB2c.2711G&amp;gt;A37yoER+yesNoCHEK2c.319+2T&amp;gt;APR+HER2-BRCA2c.8960del32yoER+ PR+ HER2-NoNo Citation Format: Thais Megid, Maria Isabel Achatz, Janina Pontes Pisani, Mariana Cartaxo. Double heterozygous pathogenic variants prevalence in a cohort of hereditary 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 PS16-12.

Abstract PS17-52: A preclinical platform of breast cancer PDX-derived cell lines as a tool for pharmacological screening and functional studies

Abstract Despite considerable progress in understanding the biology and genetics of breast cancer progression, the development of effective therapies need physiological and predictive preclinical models. In this context, breast cancer patient-derived xenograft (PDX) models has become a standard tool as they reproduce accurately the behavior of tumor of origin, in term of histological and molecular phenotype and response to chemotherapy. Although PDXs in vivo models are indispensable for preclinical studies, they suffer from some limitations due to study costs related to tumor maintenance on mice, variable engraftment rate, growth delay and limited throughput for large-scale drug screening.To address this problem and propose a time and cost effective preclinical screening tool, we developed a panel of breast cancer PDX-derived low-passage 2D cell lines as a convenient in vitro pre-screening platform to profile compound activity.30 different breast cancer PDX models including TNBC, HER2+ and ER+ were tested for their capacity to generate cell lines maintaining the characteristics of the parental PDX tumor and usable for in vitro assays.Today, we succeeded with a series of 14 PDX models.Tumor cells isolated from PDX tumor tissue were cultured under different media and matrix conditions, allowing at least 5 passages in culture. A Short Tandem Repeat (STR) comparison profile was done with the parental PDX before performing a master bank. We succeeded in establishing a panel of 14 PDX-derived cellular models (14/30 = 46% success rate).We performed short term 2D cytotoxicity assays and long term colony assays to compare cell lines in vitro drug sensitivity with their parental PDX in vivo drug response and overall, the results show that this panel reproduced the drug response profile of the original PDXs with chemotherapies, PARP inhibitors, an ADC (T-DM1) therapies.Moreover, cellular models engrafted back onto mice showed in vivo response to chemotherapies similar to that of the parental PDX confirming the identical behavior of cell line / PDX couples.As the use of cellular models is still considered as a standard for early preclinical test to evaluate drug response before moving to in vivo assays, our breast cancer PDX-derived cell line platform appeared to be a robust and relevant tool. Furthermore, since the main concern when using in vitro models is the representativeness of the results obtained when transposed to in vivo models, the similarities between cell lines and parental PDX should maximize success of further in vivo preclinical drug development. Citation Format: Stefano Cairo, Olivier Deas, Sophie Banis, Kathleen Flosseau, Enora Le Ven, Jean-Gabriel Judde. A preclinical platform of breast cancer PDX-derived cell lines as a tool for pharmacological screening and functional studies [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 PS17-52.

Treatment rationale for coronary heart disease in advanced CKD.

Chronic kidney disease (CKD) is accompanied by coronary artery disease (CAD) in most patients. In this article we describe differences in the pathogenesis, diagnosis, and treatment of CAD compared with patients without kidney impairment. The histological phenotype as well as the clinical presentation of acute and chronic coronary syndromes differ from those of patients with normal kidney function. The risk of cardiovascular events including death is strikingly increased with higher stages of CKD. Traditional but even more nontraditional cardiovascular risk factors are contributing to this increase. Screening and diagnostic procedures show limited sensitivity and specificity. Lifestyle modification is important for reducing the progression of both CKD and CAD. Aspecial emphasis should be placed on physical exercising. Equally important is astrict antihypertensive therapy due to the very high incidences of hypertension in CKD patients. Blockade of the renin-angiotensin-system is imperative providing that adverse effects can be managed. Target blood pressure should be at 130 mm Hg systolic. Antiglycemic treatment should be implemented with metformin and SGLT2-inhibitors as first-line therapy, and glomerular filtration rate thresholds must be respected for both drugs. The risk of hypoglycemia is increased with worsening kidney function. Statins are indicated for up to stage5 CKD. When arevascularization procedure is indicated (percutaneous intervention or bypass grafting), higher rates or peri-interventional morbidity and mortality must be anticipated. Taken together, the available literature on patients with CKD and CAD is clearly restricted compared with that on CAD patients with preserved kidney function. Mechanisms of arteriosclerosis and atheromatosis in CKD deserve more attention in the future. One major innovation in the field is SGLT2-inhibitor treatment with its concordant advantages for kidney and cardiac protection.

GABAA Receptors and Maternally Derived Taurine Regulate the Temporal Specification of Progenitors of Excitatory Glutamatergic Neurons in the Mouse Developing Cortex.

Temporal specification of the neural progenitors (NPs) producing excitatory glutamatergic neurons is essential for histogenesis of the cerebral cortex. Neuroepithelial cells, the primary NPs, transit to radial glia (RG). To coincide with the transition, NPs start to differentiate into neurons, undergoing a switch from symmetric to asymmetric cell division. After the onset of neurogenesis, NPs produce layer-specific neurons in a defined order with precise timing. Here, we show that GABAA receptors (GABAARs) and taurine are involved in this regulatory mechanism. Foetal exposure to GABAAR-antagonists suppressed the transition to RG, switch to asymmetric division, and differentiation into upper-layer neurons. Foetal exposure to GABAAR-agonists caused the opposite effects. Mammalian foetuses are dependent on taurine derived from the mothers. GABA and taurine function as endogenous ligands for GABAARs. Ca2+ imaging showed that NPs principally responded to taurine but not GABA before E13. The histological phenotypes of the taurine transporter knockout mice resembled those of the mice foetally exposed to GABAAR-antagonists. Foetal exposure to GABAAR-modulators resulted in considerable alterations in offspring behavior like core symptoms of autism. These results show that taurine regulates the temporal specification of NPs and that disrupting the taurine-receptor interaction possibly leads to neurodevelopmental disorders.

Nrf2 modulated the restriction of lung function via impairment of intrinsic autophagy upon real-ambient PM2.5 exposure

Compelling studies approve that fine particle matter (PM2.5) exposure was associated with high risk of respiratory disorders. However, the available data assessing the detailed influence of PM2.5 on lung was limited. To overcome the difficulty of inhalational PM2.5 exposure, the real-ambient PM2.5 exposure system was constructed. The mice were exposed to filtered air (FA) or real-ambient PM2.5 (PM2.5), and the adverse effect on lung was determined. Nuclear factor E2-related factor 2 (Nrf2) as a transcription factor, was reported to affect autophagy. Autophagy was proposed as a two-edge sword in respiratory disorders. Here, our data presented that PM2.5 exposure dramatically reduced the lung function of WT mice rather than Nrf2-/- mice. Consistently, thickened alveolar walls was observed in WT mice in PM2.5 exposure group, whereas the histological phenotype of Nrf2-/- mice exhibited no obvious alteration. Furthermore, PM2.5 exposure triggered low-grade production of inflammatory profile in WT and Nrf2-/- mice. Moreover, the protein levels of p62, Beclin1 and LC3B of WT mice rather than Nrf2-/- mice were also altered in PM2.5 exposure group. Taken together, the present study applied the real-ambient exposure system, revealed the adverse effect of air pollution on lung, and proposed the underlying mechanism.

Pre-ampullary location and size ≥10mm are independent predictors for high-grade superficial non-ampullary duodenal epithelial tumors.

The incidence of identified superficial non-ampullary duodenal epithelial tumors (SNADETs) has been increasing with recent advances in endoscopic diagnosis. Nevertheless, the clinical features of SNADETs with malignant potential remain unclear. The aim of the present study was to clarify the clinical characteristics of high-grade SNADETs. A total of 328 SNADETs that had been endoscopically or surgically resected between January 2013 and April 2019 were identified from an endoscopic and pathological database. Clinical characteristics were compared between mucosal low-grade neoplasm (n=154) and mucosal high-grade neoplasm/submucosal carcinoma (HGN/SMC, n=174). In univariate analysis, tumor size, pre-ampullary tumor location, red color, and rough/nodular surface were significantly associated with HGN/SMC. In multivariate analysis (odds ratio [95% confidence interval]), large (≥10mm) tumor size (odds ratio: 4.5, 95% confidence interval: 2.6-7.7, P<0.001) and pre-ampullary tumor location (odds ratio: 2.1, 95% confidence interval: 1.3-3.5, P=0.004) were independent predictors for HGN/SMC. Analysis of histological phenotypes revealed that the proportion of tumors that were HGN/SMC was much greater for gastric-type tumors (21/23 lesions, 91%) than for intestinal-type tumors (150/302 lesions, 50%) (P<0.001) and that all gastric-type tumors were located in the pre-ampullary portion. Pre-ampullary location and large tumor size are independent predictors for HGN/SMC SNADETs. Pre-ampullary tumor location is significantly associated with gastric histological phenotype. These findings may help in decision making for endoscopic treatment, active indication for pre-ampullary tumor or tumor ≥10mm, and understanding the pathophysiology of SNADETs.

Histologic Analysis of Testes from Prepubertal Patients Treated with Chemotherapy Associates Impaired Germ Cell Counts with Cumulative Doses of Cyclophosphamide, Ifosfamide, Cytarabine, and Asparaginase.

Cryopreservation of immature testicular tissue is an experimental strategy for the preservation of fertility in prepubertal boys that will be subjected to a gonadotoxic onset, as is the case of oncologic patients. Therefore, the objective of this study was to assess the impact of chemotherapeutic treatments on the testicular histologic phenotype in prepubertal patients. A total of 56 testicular tissue samples from pediatric patients between 0 and 16years old (28 with at least one previous chemotherapeutic onset and 28 untreated controls) were histologically analyzed and age-matched compared. At least two 5-μm sections from testis per patient separated by a distance of 100μm were immunostained for the germ cell marker VASA, the spermatogonial markers UTF1, PLZF, UCHL1, and SALL4, the marker for proliferative cells KI67, and the Sertoli cell marker SOX9. The percentage of tubule cross-sections positive for each marker and the number of positive cells per tubule cross-section were determined and association with the cumulative dose received of each chemotherapeutic drug was statistically assessed. Results indicated that alkylating agents, cyclophosphamide and ifosfamide, but also the antimetabolite cytarabine and asparaginase were associated with a decreased percentage of positive tubules and a lower number of positive cells per tubule for the analyzed markers. Our results provide new evidences of the potential of chemotherapeutic agents previously considered to have low gonadotoxic effects such as cytarabine and asparaginase to trigger a severe testicular phenotype, hampering the potential success of future fertility restoration in experimental programs of fertility preservation in prepubertal boys.