Hsa-miR-21-3p associates with breast cancer patient survival and targets genes in tumor suppressive pathways

Breast cancer is the cancer most often diagnosed in women. MicroRNAs (MIRs) are short RNA molecules that bind mRNA resulting in their downregulation. MIR21 has been shown to be an oncomiR in most cancer types, including breast cancer. Most of the effects of miR-21 have been attributed to hsa-miR-21-5p that is transcribed from the leading strand of MIR21, but hsa-miR-21-3p (miR-21-3p), transcribed from the lagging strand, is much less studied. The aim of the study is to analyze whether expression of miR-21-3p is prognostic for breast cancer. MiR-21-3p association with survival, clinical and pathological characteristics was analyzed in a large breast cancer cohort and validated in three separate cohorts, including TCGA and METABRIC. Analytical tools were also used to infer miR-21-3p function and to identify potential target genes and functional pathways. The results showed that in the exploration cohort, high miR-21-3p levels associated with shorter survival and lymph node positivity. In the three validation cohorts, high miR-21-3p levels associated with pathological characteristics that predict worse prognosis. Specifically, in the largest validation cohort, METABRIC (n = 1174), high miR-21-3p levels associated with large tumors, a high grade, lymph node and HER2 positivity, and shorter breast-cancer-specific survival (HR = 1.38, CI 1.13–1.68). This association remained significant after adjusting for confounding factors. The genes with expression levels that correlated with miR-21-3p were enriched in particular pathways, including the epithelial-to-mesenchymal transition and proliferation. Among the most significantly downregulated targets were MAT2A and the tumor suppressive genes STARD13 and ZNF132. The results from this study emphasize that both 3p- and 5p-arms from a MIR warrant independent study. The data show that miR-21-3p overexpression in breast tumors is a marker of worse breast cancer progression and it affects genes in pathways that drive breast cancer by down-regulating tumor suppressor genes. The results suggest miR-21-3p as a potential biomarker.

1331 An Observational Study Investigating the Role of Therapeutic Mammoplasty in Breast Tumours Greater Than 40mm

Aim For large breast tumours, therapeutic mammoplasty (TM) provides a breast-conserving approach to the conventional mastectomy. The prevalence and outcomes following TM in larger breast tumours is relatively unknown. This study aims to analyse the short-term outcomes and local recurrence rate following TM for breast tumours of varying sizes. Method Single-centre retrospective analysis of data from all patients undergoing a TM between June 2016-October 2019. Variables reviewed included age, imaging, tumour size, pre-operative histology, adjuvant chemotherapy and radiotherapy, post-operative pathology, post-operative complications, and recurrence rates. Results 192 patients undergoing a TM procedure were included, 126 (66%) patients had tumours <40mm and 66 (34%) patients had tumours >40mm. The average age of participants was 61 years with a mean follow-up of 31 months. The mean size of tumours >40mm was 56.8mm, of these patients 15% had positive margins, 2 (3%) patients required a further mastectomy and 8 (12%) underwent margin re-excisions. In lesions >40mm there were six episodes (9%) of T-junction delayed wound healing with two requiring surgical management, two episodes (3%) of wound infections requiring antibiotics and four episodes (6%) of seroma with one requiring surgery for an infected seroma. Two patients were found to have metastatic disease and no patients were found to have local recurrence at the most recent follow-up. Conclusions Our study demonstrated TM offers a surgical option with suitable cosmetic and oncological outcomes for women with early breast tumours above 40mm. These results warrant further study into the long-term outcomes for patients undergoing TM with tumours >40mm.

Metaplastic Breast Cancer with Chondroid Differentiation

Background: Metaplastic breast cancer (MBC) cancer is a rare subtype of breast carcinoma and carries a worse prognosis. Chondroid differentiation is the rarest among all its histologic subtypes. We report a case of MBC with chondroid differentiation and review its clinicopathological details, genetic basis, and management.Case presentation: A 56-year female presented with right-sided large breast lump. She noticed this lump 4 months before presenting. Trucut biopsy was suggestive of invasive ductal carcinoma. She underwent breast conservation surgery and histology was consistent with MBC with chondroid differentiation, pT2N3aM0. Tumour was triple-negative for ER, PR, and Her-2- neu receptors. Adjuvant treatment with chemotherapy followed by radiotherapy was given and she has been doing fine during 11 months of follow-up.Conclusion: The MBC is an uncommon subtype with heterogeneity in biological and morphological features and its knowledge is paramount while evaluating a breast lump. Understanding the pathologic and molecular basis is imperative in developing the targeted therapy to improve outcomes.

Pharmacological Assessment of the Antiprotozoal Activity, Cytotoxicity and Genotoxicity of Medicinal Plants Used in the Treatment of Malaria in the Greater Mpigi Region in Uganda

We investigated the potential antimalarial and toxicological effects of 16 medicinal plants frequently used by traditional healers to treat malaria, fever, and related disorders in the Greater Mpigi region in Uganda. Species studied were Albizia coriaria, Cassine buchananii, Combretum molle, Erythrina abyssinica, Ficus saussureana, Harungana madagascariensis, Leucas calostachys, Microgramma lycopodioides, Morella kandtiana, Plectranthus hadiensis, Securidaca longipedunculata, Sesamum calycinum subsp. angustifolium, Solanum aculeastrum, Toddalia asiatica, Warburgia ugandensis, and Zanthoxylum chalybeum. In addition, the traditional healers indicated that P. hadiensis is used as a ritual plant to boost fertility and prepare young women and teenagers for motherhood in some Ugandan communities where a high incidence of rapidly growing large breast masses in young female patients was observed (not necessarily breast cancer). We present results from various in vitro experiments performed with 56 different plant extracts, namely, 1) an initial assessment of the 16 species regarding their traditional use in the treatment of malaria by identifying promising plant extract candidates using a heme biocrystallization inhibition library screen; 2) follow-up investigations of antiprotozoal effects of the most bioactive crude extracts against chloroquine-resistant P. falciparum K1; 3) a cytotoxicity counterscreen against human MRC-5SV2 lung fibroblasts; 4) a genotoxicity evaluation of the extract library without and with metabolic bioactivation with human S9 liver fraction; and 5) an assessment of the mutagenicity of the ritual plant P. hadiensis. A total of seven extracts from five plant species were selected for antiplasmodial follow-up investigations based on their hemozoin formation inhibition activity in the heme biocrystallization assay. Among other extracts, an ethyl acetate extract of L. calostachys leaves exhibited antiplasmodial activity against P. falciparum K1 (IC50 value: 5.7 µg/ml), which was further characterized with a selectivity index of 2.6 (CC50 value: 14.7 µg/ml). The experiments for assessment of potential procarcinogenic properties of plant extracts via evaluation of in vitro mutagenicity and genotoxicity indicated that few extracts cause mutations. The species T. asiatica showed the most significant genotoxic effects on both bacterial test strains (without metabolic bioactivation at a concentration of 500 µg/plate). However, none of the mutagenic extracts from the experiments without metabolic bioactivation retained their genotoxic activity after metabolic bioactivation of the plant extract library through pre-incubation with human S9 liver fraction. While this study did not show that P. hadiensis has genotoxic properties, it did provide early stage support for the therapeutic use of the medicinal plants from the Greater Mpigi region.

Rare Copy Number Variants (CNVs) and Breast Cancer Risk

BackgroundCopy number variants (CNVs) are pervasive in the human genome but potential disease associations with rare CNVs have not been comprehensively assessed in large datasets. We analysed rare CNVs in genes and non-coding regions for 86,788 breast cancer cases and 76,122 controls of European ancestry with genome-wide array data.ResultsGene burden tests detected the strongest association for deletions in BRCA1 (P= 3.7E-18). Nine other genes were associated with a p-value < 0.01 including known susceptibility genes CHEK2 (P= 0.0008), ATM (P= 0.002) and BRCA2 (P= 0.008). Outside the known genes we detected associations with p-values < 0.001 for either overall or subtype-specific breast cancer at nine deletion regions and four duplication regions. Three of the deletion regions were in established common susceptibility loci.ConclusionsThis is the first genome-wide analysis of rare CNVs in a large breast cancer case-control dataset. We detected associations with exonic deletions in established breast cancer susceptibility genes. We also detected suggestive associations with non-coding CNVs in known and novel loci with large effects sizes. Larger sample sizes will be required to reach robust levels of statistical significance.

Rare copy number variants (CNVs) and breast cancer risk

Background: Copy number variants (CNVs) are pervasive in the human genome but potential disease associations with rare CNVs have not been comprehensively assessed in large datasets. We analysed rare CNVs in genes and non-coding regions for 86,788 breast cancer cases and 76,122 controls of European ancestry with genome-wide array data. Results: Gene burden tests detected the strongest association for deletions in BRCA1 (P= 3.7E-18). Nine other genes were associated with a p-value < 0.01 including known susceptibility genes CHEK2 (P= 0.0008), ATM (P= 0.002) and BRCA2 (P= 0.008). Outside the known genes we detected associations with p-values < 0.001 for either overall or subtype-specific breast cancer at nine deletion regions and four duplication regions. Three of the deletion regions were in established common susceptibility loci. Conclusions: This is the first genome-wide analysis of rare CNVs in a large breast cancer case-control dataset. We detected associations with exonic deletions in established breast cancer susceptibility genes. We also detected suggestive associations with non-coding CNVs in known and novel loci with large effects sizes. Larger sample sizes will be required to reach robust levels of statistical significance.