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The centrosome in animal cells is instrumental in spindle pole formation, nucleation, proper alignment of microtubules during cell division, and distribution of chromosomes in each daughter cell. Centrosome amplification involving structural and numerical abnormalities in the centrosome can cause chromosomal instability and dysregulation of the cell cycle, leading to cancer development and metastasis. However, disturbances caused by centrosome amplification can also limit cancer cell survival by activating mitotic checkpoints and promoting mitotic catastrophe. As a smart escape, cancer cells cluster their surplus of centrosomes into pseudo-bipolar spindles and progress through the cell cycle. This phenomenon, known as centrosome clustering (CC), involves many proteins and has garnered considerable attention as a specific cancer cell-targeting weapon. The kinesin-14 motor protein KIFC1 is a minus end-directed motor protein that is involved in CC. Because KIFC1 is upregulated in various cancers and modulates oncogenic signaling cascades, it has emerged as a potential chemotherapeutic target. Many molecules have been identified as KIFC1 inhibitors because of their centrosome declustering activity in cancer cells. Despite the ever-increasing literature in this field, there have been few efforts to review the progress. The current review aims to collate and present an in-depth analysis of known KIFC1 inhibitors and their biological activities. Additionally, we present computational docking data of putative KIFC1 inhibitors with their binding sites and binding affinities. This first-of-kind comparative analysis involving experimental biology, chemistry, and computational docking of different KIFC1 inhibitors may help guide decision-making in the selection and design of potent inhibitors.

Background Recent research highlights how the COVID-19 pandemic has significantly impacted alcohol consumption patterns, yet research thus far has largely overlooked the experience in West Africa. Research also has not addressed how the COVID-19 pandemic has affected access to alcohol treatment, support, and alcohol harm prevention. This study addresses this research gap in West Africa, a low-resource setting with a very high burden of alcohol harm. Objectives To understand the impact of the COVID-19 pandemic on alcohol use, access to alcohol, treatment access, and alcohol harm prevention activities in West Africa. Methods This study analyzed data from a cross-sectional online survey conducted in August and September of 2020 and distributed by the West Africa Alcohol Policy Alliance to their member alliances and stakeholders across nine countries (N = 140 participants) to understand their perceptions on COVID-19 and alcohol-related topics. Results Our findings convey a significant adverse impact on alcohol-focused NGOs and community-based organizations in West Africa. Overall, 94% of participants indicated that the COVID-19 pandemic adversely impacted their organizations’ work. In addition, 71% of participants reported reduced access to alcohol treatment or support in their communities. Lastly, 44% of the respondents indicated that people in their community drank less alcohol than usual, and only 33% answered that they perceived it to be harder to get alcohol. Conclusions These data underscore the significant impact of the COVID-19 pandemic across West Africa with respect to accessing alcohol treatment and organizational capacity to address alcohol harm. With the lack of infrastructure to address alcohol harm, this impact could exacerbate the high level of alcohol use and harm in the region.

Supplementary Figure from Hypoxia Drives Centrosome Amplification in Cancer Cells via HIF1α-dependent Induction of Polo-Like Kinase 4

This is a revised supplementary material file with unmarked changes.

Purpose: Most currently available chemotherapeutic agents target rampant cell division in cancer cells, thereby affecting rapidly dividing normal cells resulting in toxic side-effects. This nonspecificity necessitates identification of novel cellular pathways that are reprogrammed selectively in cancer cells and can be exploited to develop pharmacologically superior and less toxic therapeutics. Despite growing awareness on dysregulation of lipid metabolism in cancer cells, targeting lipid biosynthesis is still largely uncharted territory. Herein, we report development of a novel nontoxic orally deliverable anticancer formulation of monoethanolamine (Etn) for prostate cancer by targeting the Kennedy pathway of phosphatidylethanolamine (PE) lipid biosynthesis.Experimental Design: We first evaluated gastrointestinal tract stability, drug–drug interaction liability, pharmacokinetic, and toxicokinetic properties of Etn to evaluate its suitability as a nontoxic orally deliverable agent. We next performed in vitro and in vivo experiments to investigate efficacy and mechanism of action.Results: Our data demonstrate that Etn exhibits excellent bioavailability, gastrointestinal tract stability, and no drug–drug interaction liability. Remarkably, orally fed Etn inhibited tumor growth in four weeks by approximately 67% in mice bearing human prostate cancer PC-3 xenografts without any apparent toxicity. Mechanistically, Etn exploits selective overexpression of choline kinase in cancer cells, resulting in accumulation of phosphoethanolamine (PhosE), accompanied by downregulation of HIF-1α that induces metabolic stress culminating into cell death.Conclusions: Our study provides first evidence for the superior anticancer activity of Etn, a simple lipid precursor formulation, whose nontoxicity conforms to FDA-approved standards, compelling its clinical development for prostate cancer management. Clin Cancer Res; 23(14); 3781–93. ©2017 AACR.

Purpose:The purpose of this study is to predict risk of local recurrence (LR) in ductal carcinoma in situ (DCIS) with a new visualization and quantification approach using centrosome amplification (CA), a cancer cell–specific trait widely associated with aggressiveness.Experimental Design:This first-of-its-kind methodology evaluates the severity and frequency of numerical and structural CA present within DCIS and assigns a quantitative centrosomal amplification score (CAS) to each sample. Analyses were performed in a discovery cohort (DC, n = 133) and a validation cohort (VC, n = 119).Results:DCIS cases with LR exhibited significantly higher CAS than recurrence-free cases. Higher CAS was associated with a greater risk of developing LR (HR, 6.3 and 4.8 for DC and VC, respectively; P < 0.001). CAS remained an independent predictor of relapse-free survival (HR, 7.4 and 4.5 for DC and VC, respectively; P < 0.001) even after accounting for potentially confounding factors [grade, age, comedo necrosis, and radiotherapy (RT)]. Patient stratification using CAS (P < 0.0001) was superior to that by Van Nuys Prognostic Index (VNPI; HR for CAS = 6.2 vs. HR for VNPI = 1.1). Among patients treated with breast-conserving surgery alone, CAS identified patients likely to benefit from adjuvant RT.Conclusions:CAS predicted 10-year LR risk for patients who underwent surgical management alone and identified patients who may be at low risk of recurrence, and for whom adjuvant RT may not be required. CAS demonstrated the highest concordance among the known prognostic models such as VNPI and clinicopathologic variables such as grade, age, and comedo necrosis.

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Background: Triple-negative breast cancer (TNBC) comprises 10%-20% of invasive breast carcinoma cases. Patients with TNBC exhibit poorer 5-year survival and higher rates of metastasis and recurrence than patients with other breast cancer subtypes. TNBC is further divided based on gene expression profiling into androgen receptor (AR)-positive (AR+) TNBC and AR-negative TNBC (or quadruple-negative breast cancer [QNBC]). While AR is an emerging therapeutic target for AR+ TNBC, there are no treatment options for QNBC. AR expression is associated with favorable disease-free, overall, and recurrence-free survival in patients with TNBC, suggesting that AR loss in TNBC tumors confers an aggressive disease course. Genomic instability has been recognized as one of the drivers of tumorigenesis and can be evidenced by the presence of chromosomal instability, such as DNA copy number alterations (CNAs). Although there are many studies characterizing the genomic profiles of TNBCs, little is known about the tumor biology and genetic makeup of QNBCs. In this study, we explored the impact of CNAs on microRNA (miRNA) expression and, subsequently, on signaling pathways associated with QNBC aggressiveness and clinical outcomes. Methods: AR expression was immunohistochemically assessed in tissues of 53 patients with TNBC from the Histopathology and Tissue Shared Resources of Georgetown University. AR positivity was defined as AR expression in >1% of cells. Genome-wide copy number profiling of formalin-fixed paraffin-embedded samples was performed using the Agilent SurePrint G3 Human CGH Microarray, and miRNA expression analysis was conducted using NanoString nCounter Human v3a miRNA Expression Assay. Differentially expressed miRNAs between TNBC and. QNBC samples were integrated with array CGH data from the same tissue sample to identify common gene targets that may be affected by both CNAs and miRNA deregulation. Functional enrichment analysis was performed to identify cancer pathways deregulated in QNBC. The expression levels of Centrosome Amplification 20 (CA20) and Chromosome Instability 25 (CIN 25) gene signatures were evaluated in 524 primary invasive breast cancers from The Cancer Genome Atlas (TCGA) database. Results: We observed that 64% of TNBC samples lacked AR expression and that QNBCs exhibited a significantly higher CNA level (p50% of QNBCs. Additionally, CNAs affecting genes of the CIN25 signature were significantly higher in QNBC than in TNBC; however, CNAs in CA20 signature genes were similar in QNBC and TNBC samples. Notably, the expression levels of CA20 and CIN25 genes were significantly higher in QNBC than in TNBC. Moreover, we identified 184 differentially expressed miRNAs between QNBC and TNBC samples (p Citation Format: Shristi Bhattarai, Bruna M. Sugita, Luciane Cavalli, Ritu Aneja. Androgen receptor loss is associated with genomic instability characterized by copy number alterations and mirna deregulation in triple-negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-09-13.

Background: Pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) isassociated with a favorable prognosis and low recurrence rate, particularly in patients with triple-negative breast cancer (TNBC), an aggressive breast cancer subtype. However, only 30%-40%of patients with TNBC show pCR after standard NAC, while the remaining patients have non-pCR or residual disease. Different histological components of the tumor, including tumor stroma,polyploid giant cancer cells, and immune cells, can influence response to NAC in patients withTNBC. However, it remains unknown which histological components contribute to pCR afterNAC in some patients with TNBC but not in others. To address this, we developed a machinelearning pipeline to identify the most significant histological components contributing to NACresponse in patients with TNBC. Different representations of histological components were usedin this pipeline as a "full-slide" feature set to predict NAC response in patients with TNBC.Methods: Post-NAC tissues from 129 patients (55 pCR and 74 non-pCR) from Emory DecaturHospital (Decatur, GA) were used. The Decatur cohort was separated based on NAC responseinto training (33 pCR and 33 non-pCR) and validation sets. There were a total of 17 histologicalclasses annotated by a board-certified pathologist and two NAC labels. Our pipeline comparedfour different machine learning approaches (1NN, ensembleTree, linSVM, and rbfSMV), and thebest classifiers were used for testing. Each whole slide image (WSI) was partitioned into smallertiles. The first classifier used texture-based features from the 17 histological classes. Inaggregation, 55 texture features derived from each tile were provided to a classifier (rbfSVM) tooutput a probability for each new tile during testing. The predicted histology labels were used togenerate a tile-level classification map that recaptured NAC response to predict pCR or non-pCR. Each such tile-level classification map was represented by 80 graph-based featurescapturing the relevant spatial information across the different histological classes. These graph-based features were provided to the best patient-level classifier (rbfSVM) to predict NACresponse for each patient.Results: Our tile-level classification achieved 83% histological classification accuracy, and ourpatient-level machine learning pipeline achieved a 69% NAC classification accuracy. The keyhistological classes that helped distinguish between pCR and non-pCR were tumor & immunecells, tumor & stroma, and microvessel density (MVD) & immune cells. Patients with thesecombinations of discriminating histological classes were 69% likely to have a pCR after NACbased on the accuracy of the rbfSVM NAC classifier. This finding suggests that a classificationmap with more pCR combinations in WSI classified by rbfSVM NAC classifier can helpdistinguish pCR from non-pCR. Notably, our pipeline required less than 24 hours to analyzeWSIs of one patient, accelerating patient selection for NAC treatment.Conclusion: Our machine learning pipeline can robustly identify the most significanthistological classes predicting NAC response and can guide patient selection for NAC treatment. Citation Format: Timothy Byron Fisher, Hongxiao Li, Rekha TS, Jayashree Krishnamurthy, Shristi Bhattarai, Emiel A.M. Janssen, Jun Kong, Ritu Aneja. Using machine learning approaches to predict response to neoadjuvant chemotherapy in patients with triple-negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-08-16.

Background: Neoadjuvant chemotherapy (NAC) is becoming the standard of care for aggressive breast cancer (BC). In patients with BC, NAC can increase eligibility for breast-conserving surgery, lower the excision volume, and assist in evaluating the disease course. NAC may shrink (residual disease, RD) or even eliminate (pathologic complete response, pCR) the tumor. Patients with RD often have a poor long-term prognosis. In contrast, pCR is associated with better long-term survival outcomes. Approximately 30% of patients with BC show pCR after NAC, while the rest exhibit RD. A critical barrier to improving NAC outcomes in patients with BC is the lack of precise molecular targets and limited understanding of the molecular mechanisms underlying differential treatment outcomes in patients with BC receiving NAC. Thus, there is an urgent clinical need to elucidate the underlying molecular regulation associated with NAC resistance. Studies have shown that cellular stress induces the release of small extracellular vesicles known as exosomes, contributing to drug resistance. In this study, we evaluated the ability of exosomal metabolic profiles to predict NAC response in BC patients. Methods: We collected blood samples of 16 patients with BC who received NAC at Grady Memorial Hospitals, USA. Further, total exosomes were isolated from plasma by an ultracentrifugation method and characterized for their concentration (number/ml), size distribution, and exosomal content. Exosome sizes were validated using nanoparticle tracking analysis. RNA, proteins, and metabolites were isolated from exosomes to perform deep multiplatform analyses (viz. miRNA seq, proteomics, and metabolomics) of exosomal cargo to identify various potential biological determinants of NAC response and evaluate their contributions to the risk of RD in patients with BC after NAC. KEGG and HMDB IDs were used for metabolomics data analyses. Various analyses viz. enriched metabolomic pathway analysis, MS peak to pathway analysis (using GSEA and Mummichog), and network explorer analysis were performed using Metaboanalyst online portal to reveal the metabolic signatures in exosomes of BC patients who exhibited pCR and RD after NAC treatment. Results: Among the 16 patients with BC who received NAC, eight patients attained pCR, and eight had RD. Interestingly, we found that patients with pCR secreted 61% fewer exosomes but had 24% larger-sized exosomes than patients with RD following NAC. Furthermore, significant differences in the loading of exosomal cargo were observed between the two groups. Based on these results, we proposed that the differential RNA, protein, and metabolite profile of exosomes might contribute to the higher drug sensitivity in patients with pCR. Our proteomics and miRNA-seq analyses of exosomes from patients with pCR and RD after NAC did not show any significant differences between the groups. However, metabolite set enrichment analysis using MetaboAnalyst revealed that compared to exosomes from patients with pCR, patients with RD were significantly enriched in various metabolic pathways. These pathways included valine, leucine, and isoleucine biosynthesis; phenylalanine metabolism; one-carbon pool folate; and inositol phosphate metabolism. Additionally, MS peak to pathway analysis revealed differential enrichment of various other metabolic pathways, particularly in patients with RD.Conclusions: These data strongly suggest that exosomal metabolic signatures are associated with differential NAC outcomes in patients with BC. We believe that in-depth studies of these metabolite signatures will give essential clues about differential NAC response in patients with BC. Citation Format: Shriya Joshi, Chakravarthy Garalapati, Shristi Bhattarai, Darshan Shimoga Chandrashekar, Sooryanarayana Varambally, Gagan Deep, Ritu Aneja. Exosomal metabolic signatures are associated with differential response to neoadjuvant chemotherapy in patients with breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P2-12-24.

Background: Triple-negative breast cancer (TNBC) is a notoriously hard to treat and aggressive breast cancer (BC) subtype. Androgen receptor-positive (AR+) TNBC is characterized by AR overexpression and sensitivity to AR antagonists. In contrast, basal-like TNBC lacking AR expression, also known as quadruple-negative breast cancer (QNBC), is refractory to AR antagonists. Women of African descent are predisposed to TNBC. Moreover, QNBC differences between African American (AA) and European American (EA) patients have been noted, with the former expressing distinct basal and immune signatures and having worse outcomes. Although AR testing is not part of standard BC diagnosis and the QNBC subtype is usually reported as TNBC, QNBC may have distinct biological and clinical characteristics. In this study, we examined the prevalence and clinical characteristics of QNBC.Methods: We immunohistochemically evaluated AR expression in formalin-fixed paraffin-embedded TNBC samples (n=760) from our multi-institutional cohorts across the US and Nigeria. Samples with ≥1% AR-positive nuclei were deemed AR-positive. Descriptive statistics were generated for clinicopathological characteristics. Kaplan-Meier (KM) survival analyses were performed to compare overall survival (OS) between patient groups with a different AR status. Multivariable Cox regression analysis was conducted to select the covariates (a≥0.20 removal criterion). AR mRNA levels in patients with BC were evaluated using RNA-seq data retrieved from The Cancer Genome Atlas (TCGA). The Cancer Proteome Atlas (TCPA) BRCA dataset was used to assess AR protein levels.Results: We found an association between African ancestry and AR loss. QNBC accounted for 92.1% of BCs in Africans, 82.4% in AAs, and only 39.8% in EAs. Similarly, analyses of RNA-seq data from TCGA revealed that tumors in AAs with TNBC (n=32) had lower AR mRNA levels than their EA counterparts (n=67). Consistently, AR protein levels in TNBC tumors were lower in AAs than in EAs. Univariate analyses revealed a significant association between QNBC status and high tumor grade (grade 3: 83.02% vs. 66.41%, p2 cm (60.88% AAs vs. 45.65% EAs, p=0.075). KM survival analyses indicated that patients with QNBC had worse outcomes than those with TNBC, with significant differences in women of African descent (p Citation Format: Shristi Bhattarai, Geetanjali Saini, Manali Rupji, Justin Luningham, Ravi C. Turaga, Uma Krishnamurti, Xiaoxian (Bill) Li, Johnson Agboola, Murtala Abubakar, Haruna A. Nggada, Abidemi Omonisi, Saad A. Ahmed, Luciane Cavalli, Ritu Aneja. Racial disparities in breast cancer chiefly reside in the lesser-known quadruple-negative breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P4-07-31.

Background: Triple-negative breast cancer (TNBC) is a heterogeneous breast cancer subtype with distinct biological features and clinical behavior. TNBC is associated with an increased risk of metastasis and recurrence. In this whole exome study, we investigated the genetic profiles of primary TNBC tumors and paired metastases using next-generation sequencing (NGS). Methods: Genomic DNA extracted from 35 paraffin-embedded formalin-fixed (FFPE) tissues (15 primary tumors that did not metastasize, 11 primary tumors that metastasized, and nine paired metastatic tumors to the lymph nodes) was analyzed by whole exome sequencing (WES). Reads were trimmed using Trim-Galore and were aligned to the reference genome GRCh38 using Burrows-Wheeler Aligner (BWA). BAM files were preprocessed to optimize variant calling, and variant calling was carried out using the GATK pipeline. Variants were annotated using SnpEff. Tumors were analyzed for single nucleotide variants (SNV), point mutations, and insertions/deletions (indels). Results: Primary tumors that did not metastasize had a higher number of variants (~13,500) than primary tumors that metastasized (~12,900). However, the number of variants was similar between the primary tumors (~12,900 variants) and their matched metastases (~12,500 variants). MUC3A was the top mutated gene both in primary tumors that did not metastasize and in those that metastasized. MUC3A was also the top mutated gene in matched metastatic lesions. Moreover, we compared the mutational status of the most frequently mutated genes in TNBC samples from the TCGA and METABRIC datasets. We found that in our dataset, TP53, MAP3K1, and PTEN were mutated in 60%, 93%, and 7% of primary TNBC tumors without metastases and in 36%, 0%, and 36% of primary TNBC tumors that metastasized. Mutations in TP53 and PTEN were found in 2/9 and 1/9 of primary and metastases pairs, respectively. No mutations in PI3KCA were observed in any of the primary or matched metastatic tumors. Conclusions: We used WES to compare the genomic landscapes of primary TNBC tumors and matched metastatic tumors, as well as of primary TNBC tumors that metastasized and those that did not metastasize. We found very similar genomic alterations between the primary and paired metastatic tumors, indicating that genomic features may be retained during metastasis. Primary tumors that did not metastasize showed a greater extent of genomic alterations than primary tumors that metastasized. Citation Format: Jaspreet Kaur, Darshan S. Chandrashekar, Zsuzsanna Varga, Emiel Janssen, Khanjan Gandhi, Karuna Mittal, Umay Kiraz, Sooryanarayana Varambally, Ritu Aneja. Whole exome sequencing of matched primary and metastatic triple-negative breast cancer samples [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-09-14.

Extensive intratumoral heterogeneity (ITH) is believed to contribute to therapeutic failure and tumor recurrence, as treatment-resistant cell clones can survive and expand. However, little is known about ITH in triple-negative breast cancer (TNBC) because of the limited number of single-cell sequencing studies on TNBC. In this study, we explored ITH in TNBC by evaluating gene expression-derived and imaging-derived multi-region differences within the same tumor. We obtained tissue specimens from 10 TNBC patients and conducted RNA sequencing analysis of 2–4 regions per tumor. We developed a novel analysis framework to dissect and characterize different types of variability: between-patients (inter-tumoral heterogeneity), between-patients across regions (inter-tumoral and region heterogeneity), and within-patient, between-regions (regional intratumoral heterogeneity). We performed a Bayesian changepoint analysis to assess and classify regional variability as low (convergent) versus high (divergent) within each patient feature (TNBC and PAM50 subtypes, immune, stroma, tumor counts and tumor infiltrating lymphocytes). Gene expression signatures were categorized into three types of variability: between-patients (108 genes), between-patients across regions (183 genes), and within-patients, between-regions (778 genes). Based on the between-patient gene signature, we identified two distinct patient clusters that differed in menopausal status. Significant intratumoral divergence was observed for PAM50 classification, tumor cell counts, and tumor-infiltrating T cell abundance. Other features examined showed a representation of both divergent and convergent results. Lymph node stage was significantly associated with divergent tumors. Our results show extensive intertumoral heterogeneity and regional ITH in gene expression and image-derived features in TNBC. Our findings also raise concerns regarding gene expression based TNBC subtyping. Future studies are warranted to elucidate the role of regional heterogeneity in TNBC as a driver of treatment resistance.

Precision (or personalized) medicine holds great promise in the treatment of breast cancer. The success of personalized medicine is contingent upon inclusivity and representation for minority groups in clinical trials. In this article, we focus on the roadblocks for the African American demographic, including the barriers to access and enrollment in breast oncology trials, the prevailing classification of race and ethnicity, and the need to refine monolithic categorization by employing genetic ancestry mapping tools for a more accurate determination of race or ethnicity.

Introduction Adrenal gland lesions span a range of entities from benign and malignant primary neoplasms to metastatic tumors. Fine-needle aspiration (FNA) provides a minimally invasive diagnostic tool to stage patients with known malignancy and procure material for molecular testing. This study characterizes the clinicopathologic associations of patients with adrenal gland FNA from 2 large hospitals. Materials and methods FNAs were identified by query of electronic medical record from 2002-2019. Clinical and pathological information was collated and correlated with corresponding surgical diagnosis when available. Results Of 139 cases, the majority (n = 127, 91%) were adequate computed tomography–guided FNAs and included the following diagnoses: positive for malignancy (n = 77, 55%), negative for malignancy (n = 32, 23%), neoplastic cells present (n = 16, 12%), nondiagnostic (n = 12, 9%), atypical (n = 1, 1%), and suspicious for carcinoma (n = 1, 1%). The majority (94%, n = 72 of 77) of malignancies were metastatic tumors, most frequently carcinoma (n = 53 of 72, 74%), followed by melanoma (n = 11 of 72, 15%), lymphoma (n = 4 of 72, 6%), and sarcoma (n = 4 of 72, 6%). Metastatic carcinomas included lung (n = 21 of 72, 29%), genitourinary (n = 12 of 72, 17%), and hepatobiliary or gastrointestinal tract (n = 11 of 72, 15%) primaries. Primary adrenal neoplasms (n = 23) included adenomas (n = 11 of 23, 48%), pheochromocytomas (n = 4 of 23, 17%), and myelolipomas (n = 3 of 23, 13%). Thirty-two patients with metastases died of disease after median follow-up of 8 months. Conclusions High specimen adequacy (n = 127, 91%) and low indeterminate rates (n = 2, 2%) are achieved with adrenal FNA. Most aspirated lesions represent metastases from primary lung carcinomas, but other primary sites including those below the diaphragm are part of the diagnostic differential. Adrenal metastasis was associated with a poor prognosis, with median survival of 8 months.

For routine pathology diagnosis and imaging based biomedical research, Whole-Slide Image (WSI) analyses have been largely limited to a 2D tissue image space. For a more definitive tissue representation to support fine-resolution spatial and integrative analyses, it is critical to extend such tissue based investigations to a 3D tissue space with spatially aligned serial tissue WSIs in different stains, such as Hematoxylin and Eosin (H&E) and Immunohistochemistry (IHC) biomarkers. However, such WSI registration is technically challenged by the overwhelming image scale, the complex histology structure change, and the significant difference in tissue appearances in different stains. We propose a novel translation based deep learning registration network CycGANRegNet that spatially aligns serial WSIs stained in H&E and by IHC biomarkers without prior deformation information for the model training. First, synthetic IHC images are produced from H&E slides through a robust image synthesis algorithm. Next, the synthetic and the real IHC images are registered through a Fully Convolutional Network with multi-scaled deformable vector fields and a joint loss optimization. We perform the registration at the full image resolution, retaining the tissue details in the results. Evaluated with a dataset of 76 breast cancer patients with one H&E and two IHC serial WSIs for each patient, CycGANRegNet outperforms multiple state-of-the-art deep learning based and conventional pathology image registration methods. Our results suggest that CycGAN-RegNet can produce promising registration results with serial WSIs in different stains, enabling integrative 3D tissue-based biomedical investigations.

Ginger (Zingiber officinale) is one of the most widely consumed dietary supplements worldwide. Its anticancer potential has been demonstrated in various studies. However, ginger roots obtained from different geographical locations showed extensive variability in their activities, mainly due to differences in the levels of bioactive compounds. Here we evaluated the effect of these differences on the anticancer activity of ginger by performing efficacy-based fingerprinting. We characterized the fingerprint profiles of 22 ginger samples using liquid chromatography-mass spectroscopy, followed by a principal component analysis (PCA) and pearson correlation analysis. We also evaluated the anti-proliferative effects (IC50) of these samples on triple-negative breast cancer cells using the MTT assays. The supervised PCA identified a subset of analytes whose abundance strongly associated with the IC50 values of the ginger extracts, providing a link between ginger extract composition and in vitro anticancer efficacy. This study demonstrated that variation in the ginger fingerprint profiles resulting from differences in their chemical composition could have a significant impact on efficacy and bioactivity of ginger extracts. Also, this first-of-a-kind efficacy-based fingerprinting approach proposed here can identify potent anticancer candidates from the ginger fingerprint without the need for isolating individual components from the extracts.

Centrosome amplification (CA) has been implicated in the progression of various cancer types. Although studies have shown that overexpression of PLK4 promotes CA, the effect of tumor microenvironment on polo-like kinase 4 (PLK4) regulation is understudied. The aim of this study was to examine the role of hypoxia in promoting CA via PLK4. We found that hypoxia induced CA via hypoxia-inducible factor-1α (HIF1α). We quantified the prevalence of CA in tumor cell lines and tissue sections from breast cancer, pancreatic ductal adenocarcinoma (PDAC), colorectal cancer, and prostate cancer and found that CA was prevalent in cells with increased HIF1α levels under normoxic conditions. HIF1α levels were correlated with the extent of CA and PLK4 expression in clinical samples. We analyzed the correlation between PLK4 and HIF1A mRNA levels in The Cancer Genome Atlas (TCGA) datasets to evaluate the role of PLK4 and HIF1α in breast cancer and PDAC prognosis. High HIF1A and PLK4 levels in patients with breast cancer and PDAC were associated with poor overall survival. We confirmed PLK4 as a transcriptional target of HIF1α and demonstrated that in PLK4 knockdown cells, hypoxia-mimicking agents did not affect CA and expression of CA-associated proteins, underscoring the necessity of PLK4 in HIF1α-related CA. To further dissect the HIF1α-PLK4 interplay, we used HIF1α-deficient cells overexpressing PLK4 and showed a significant increase in CA compared with HIF1α-deficient cells harboring wild-type PLK4. These findings suggest that HIF1α induces CA by directly upregulating PLK4 and could help us risk-stratify patients and design new therapies for CA-rich cancers. Implications: Hypoxia drives CA in cancer cells by regulating expression of PLK4, uncovering a novel HIF1α/PLK4 axis.

Our research sought to describe barriers to mammography screening among a sample of predominantly Black women in metropolitan Atlanta, Georgia.The Pink Panel project convened community leaders from faith-based institutions to administer an offline survey to women via convenience sampling at fourteen churches in Atlanta in late 2019 and early 2020. With the COVID-19 pandemic, the research team switched to an online survey. The survey included seven questions about breast cancer awareness, barriers to breast cancer screening, and screening status. We used residence information to attain the 9-digit zip code to link to the Area Deprivation Index at the Census Block Group neighborhood level. We report results as descriptive statistics of the barriers to mammography screening.The 643 women represented 21 counties in Georgia, predominantly from metropolitan Atlanta, and 86% identified as Black. Among women aged 40 and older, 90% have ever had a mammogram. Among all women, 79% have ever had a mammogram, and 86% indicated that they would get a mammogram if offered in their neighborhood. The top barriers to mammography screening were lack of health insurance and high cost. Barriers to mammography screening did not differ substantially by Area Deprivation Index.Among metropolitan Atlanta women aged 40+ , nearly all reported ever having a mammogram. However, addressing the barriers, including lack of health insurance and high cost, that women reported may further improve mammography screening rates.

MiR-150-5p is frequently deregulated in cancer, with expression and mode of action varying according to the tumor type. Here, we investigated the expression levels and role of miR-150-5p in the aggressive breast cancer subtype triple-negative breast cancer (TNBC). MiR-150-5p expression levels were analyzed in tissue samples from 113 patients with invasive breast cancer (56 TNBC and 57 non-TNBC) and 41 adjacent non-tumor tissues (ANT). Overexpression of miR-150-5p was observed in tumor tissues compared with ANT tissues and in TNBC compared with non-TNBC tissues. MiR-150-5p expression levels were significantly associated with high tumor grades and the Caucasian ethnicity. Interestingly, high miR-150-5p levels were associated with prolonged overall survival. Manipulation of miR-150-5p expression in TNBC cells modulated cell proliferation, clonogenicity, migration, and drug resistance. Manipulation of miR-150-5p expression also resulted in altered expression of its mRNA targets, including epithelial-to-mesenchymal transition markers, MYB, and members of the SRC pathway. These findings suggest that miR-150-5p is overexpressed in TNBC and contributes to the aggressiveness of TNBC cells in vitro.

Triple-negative breast cancer (TNBC) is a heterogeneous breast cancer subtype that lacks expression of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Approximately 15%-20% of breast cancers are triple-negative. Based on expression of the androgen receptor (AR), TNBC can be categorized as AR-positive (AR+ TNBC) or AR-negative (quadruple-negative breast cancer, QNBC). Studies have reported that the loss of AR expression is associated with poor prognoses. Recent evidence suggests that QNBCs disproportionately affect women of African descent and contributes to poor overall survival. QNBC is a more aggressive disease compared to TNBC, and patients with QNBC currently have no treatment options beyond chemotherapy. Data from interrogating public databases such as TCGA indicates that QNBC is distinct from TNBC in molecular makeup, particularly in immune tumor microenvironment. Currently, QNBC is not considered distinct from the broader TNBC subtype, hence a comprehensive molecular characterization is lacking. We aim to examine differences between the TNBC and QNBC tumor microenvironments that consist of both immune and stromal components. We analyzed RNA expression data from the METABRIC (TNBC= 11, QNBC= 185) and TCGA (TNBC= 29, QNBC= 103) datasets. Based on AR transcript expression, patients with TNBC were stratified into AR-low (considered QNBC) and AR-high (deemed ‘true’ TNBC) groups. The analytical immune tools - CIBERSORTx and xCELL were used to analyze the RNA-Seq data. CIBERSORTx can distinguish 22 mature human hematopoietic populations and provides abundance of member cell types in a mixed cell population, from gene expression data. xCELL is a gene signature-based method that can perform cell type enrichment analysis from gene expression for 64 immune and stromal cell types. Thus, xCell can discriminate between closely related cell types. Using these immune tools, the tumor microenvironments of the TNBC and QNBC groups were compared. Correlations between individual immune and stromal components with patient survival probability were assessed. Significant differences (p-value Citation Format: Geetanjali Saini, Manali Rupji, Mahak Bhargava, Sunil Badve, Ritu Aneja. Profiling the tumor microenvironments of triple- and quadruple-negative breast cancers. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4628.

This research had two aims: (1) to assess how often bisexual and lesbian women self-report screening and counseling for alcohol use in primary care settings; and (2) understand how bisexual and lesbian women respond to brief messages that alcohol increases breast cancer risk. The study sample consisted of 4891 adult U.S. women who responded to an online, cross-sectional Qualtrics survey in September–October 2021. The survey included the Alcohol Use Disorders Identification Test (AUDIT), questions about alcohol screening and brief counseling in primary care, and questions assessing awareness of the link between alcohol use and breast cancer. Bivariate analyses and logistic regression were conducted. Bisexual and lesbian women had higher odds of harmful drinking (AUDIT score ≥ 8) than heterosexual women (adjusted odds ratio [AOR] = 1.26, 95% confidence interval [CI] = 1.01–1.57 for bisexual women; AOR =1.78, 95% CI = 1.24–2.57 for lesbian women). However, bisexual and lesbian women were no more likely than heterosexual women to be advised about drinking in primary care. In addition, bisexual, lesbian, and heterosexual women had similar reactions to messages highlighting that alcohol is a risk factor for breast cancer. Women across all three sexual orientations who are harmful drinkers more often agreed to search for more information online or talk to a medical professional compared to non-harmful drinkers.

The abysmal success rate of anticancer drugs in clinical trials, is in part, attributable to discordance between cultured cancer cells and patient tumors. While tumors in vivo, display a lower mitotic index, patient tumors portray much higher centrosomal aberrations, relative to in vitro cultured cells. The microenvironment too differs considerably between the in vitro and in vivo scenarios. Notably, another hallmark of cancer, hypoxia, is not recapitulated in cell lines cultured under normoxic conditions. These observations raise the possibility that hypoxia may be the missing link that explains the discordance between cell biological phenomena in vitro versus physiological conditions. Further, the interplay between hypoxia and centrosome amplification (CA) is relatively understudied. Recent research from our laboratory, geared toward examining the biological link between the two, has uncovered that hypoxia induces the expression of proteins (Plk4, Aurora A, Cyclin D) implicated in CA, in a hypoxia-inducible factor 1α (HIF-1α)-dependent context. Our studies evidence that hypoxia fuels CA that underlie intratumoral heterogeneity and metastatic potential of cancer cells. Given the advent of HIF-1α inhibitors, this research has ramifications in aiding patient risk stratification and designing new cancer drug therapies to facilitate clinical decision-making.