Purpose The recent development of multi-gene assays for gene expression profiling has contributed significantly to the understanding of the clinically and biologically heterogeneous breast cancer (BC) disease. PAM50 is one of these assays used to stratify BC patients and individualize treatment. The present study was conducted to characterize PAM50-based intrinsic subtypes among Ethiopian BC patients. Patients and methods Formalin-fixed paraffin-embedded tissues were collected from 334 BC patients who attended five different Ethiopian health facilities. All samples were assessed using the PAM50 algorithm for intrinsic subtyping. Results The tumor samples were classified into PAM50 intrinsic subtypes as follows: 104 samples (31.1%) were luminal A, 91 samples (27.2%) were luminal B, 62 samples (18.6%) were HER2-enriched and 77 samples (23.1%) were basal-like. The intrinsic subtypes were found to be associated with clinical and histopathological parameters such as steroid hormone receptor status, HER2 status, Ki-67 proliferation index and tumor differentiation, but not with age, tumor size or histological type. An immunohistochemistry-based classification of tumors (IHC groups) was found to correlate with intrinsic subtypes. Conclusion The distribution of the intrinsic subtypes confirms previous immunohistochemistry-based studies from Ethiopia showing potentially endocrine-sensitive tumors in more than half of the patients. Health workers in primary or secondary level health care facilities can be trained to offer endocrine therapy to improve breast cancer care. Additionally, the findings indicate that PAM50-based classification offers a robust method for the molecular classification of tumors in the Ethiopian context.

Background The operating room (OR) is one of the most expensive areas of a hospital, requiring large capital and recurring investments, and necessitating efficient throughput to reduce costs per patient encounter. On top of increasing costs, inefficient utilization of operating rooms results in prolonged waiting lists, high rate of cancellation, frustration of OR personnel as well as increased anxiety that negatively impacts the health of patients. This problem is magnified in developing countries, where there is a high unmet surgical need. However, no system currently exists to assess operating room utilization in Ethiopia. Methodology A prospective study was conducted over a period of 3 months (May 1 to July 31, 2019) in a tertiary hospital. Surgical case start time, end time, room turnover time, cancellations and reason for cancellation were observed to evaluate the efficiency of eight operating rooms. Results A total of 933 elective procedures were observed during the study period. Of these, 246 were cancelled, yielding a cancellation rate of 35.8%. The most common reasons for cancellation were related to lack of OR time and patient preparation (8.7% and 7.7% respectively). Shortage of facilities (instrument, blood, ICU bed) were causes of cancelation in 7.7%. Start time was delayed in 93.4% (mean 8:56 am ± 52 min) of cases. Last case completion time was early in 47.9% and delayed in 20.6% (mean 2:54 pm ± 156 min). Turnover time was prolonged in 34.5% (mean 25 min ± 49 min). Total operating room utilization ranged from 10.5% to 174%. Operating rooms were underutilized in 42.7% while overutilization was found in 14.6%. Conclusion We found a high cancellation rate, most attributable to late start times leading to delays for the remainder of cases, and lack of preoperative patient preparation. In a setting with a high unmet burden of surgical disease, OR efficiency must be maximized with improved patient evaluation workflows, adequate OR staffing and commitment to punctual start times. We recommend future quality improvement projects focusing on these areas to increase OR efficiency.

Background: Breast cancer is the most common cancer overall, with an estimated 2.4 million incident cases in 2015. Current treatment of breast cancer is mainly surgery, chemotherapy, radiotherapy, and hormonal therapy. Although these approaches appear to be effective, they have failed to result in long-term cure, especially for aggressive HER2-positive and triple-negative breast cancer. This particularly holds for sub-Saharan Africa with a dominance of aggressive breast cancer subtypes with very limited treatment options and almost no research activities in place. As a result, continuous exploration of alternative treatment strategies is urgently required. Consequently, a collaborative research program has been established between Addis Ababa University, Ethiopia, and Martin-Luther-University, Germany, to enhance breast cancer research in Ethiopia. As a pilot study in Germany, we have assessed the in vitro potency of the three inhibitors romidepsin, trametinib, and lapatinib targeting HDAC, MEK and HER2/EGFR, respectively, in different breast cancer cell lines. Methods: Seven distinct breast cancer cell lines (MCF7, SKBR3, T47D, BT474, BT20, HCC1806 and HCC1937) of known HER-2 and hormone status were used to evaluate the antiproliferative effect of romidepsin, lapatinib, and trametinib. Cells cultivated in complete RPMI media were seeded in to 96 well plates (3,000 cells/well) and incubated at 370C and 5% CO2 for 24hrs. The cells were then treated for 24, 48, and 72 hours with different concentrations of the drugs. Cells exposed to solvent alone served as controls. XTT cell viability assay was used to determine proliferation of cells following manufacturers' instruction. IC50 value was calculated (Compusyn software) for each drug against the different cell lines to determine the antiproliferative effect. Results: Romidepsin exhibited a strong antiproliferative effect against all breast cancer cell lines tested, resulting in a marked decrease in cell viability with increasing concentrations, which highly varied between the breast cancer cell lines analyzed. The calculated IC50 values 48 hrs after the onset of treatment ranged between 2.2 and 52 nM; T47D was the most sensitive. Lapatinib treatment of the EGFR/HER-2 expressing BT20, SKBR3 and BT474 caused dose-dependent suppression of cell viability against SKBR3 and BT474 with an IC50 value of 450 and 77 nM, respectively. In contrast, BT20 cells were resistant to this treatment and were the only breast cancer cell line with an enhanced sensitivity to increasing concentrations of trametinib (IC50 =2.79 μM). Conclusion: In sum, romidepsin might improve the treatment efficacy of breast cancer though it needs to be further explored in vitro as well as in vivo alone and in combination with other therapies. Although trametinib has been suggested to be insufficient for monotherapy of breast cancer in general, it might be still a candidate for the treatment of basal-like breast cancers due to the sensitivity of BT20 cells. Citation Format: Meron Yohannes Nigussie, Jürgen Bukur, Zelalem Desalegn Woldesonbet, Tamrat Abebe Zeleke, Yonas Bekuretsion, Mahlet Arayselassie, Mathewos Assefa, Abebe Bekele, Endale Anberber, Martina Vetter, Claudia Wickenhauser, Eva J Kantelhardt, Barbara Seliger. Antiproliferative efficacy of different targeted drugs on breast cancer cell lines of distinct subtypes [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr B020.

Background: Breast cancer is becoming a major public health problem in developing countries. Accumulated evidence noted that the majority of breast cancer patients are in a younger age group. Moreover, most breast cancer patients appear in health institutions at late stages of the disease, which compromises the clinical outcome and disease management. With regard breast cancer, it solely depends on pathologic and clinical diagnoses. The characterization of hormone receptors and HER-2 status using immunohistochemistry (IHC) has been introduced in Ethiopia; however, there is critical problem with the provision of requirements for the test. As a result, the development and establishment of a rapid, reproducible, and alternative diagnostic method in resource limited setting is essential. The current research work aims to optimize endpoint PCR as a tool for determination of ESR1, PGR, and ERBB2 expression level using breast cancer specimen in Ethiopian patients. Furthermore, the project work will be extended to implement the method into clinical practice in the Ethiopian settings. Methods: The experiments were started with known ESR1, PGR, and ERBB2 RNA expression in different breast cancer cell lines such as MCF-7, MDA-MB-231, BT-20, and SKBR-3 .cDNA from the breast cancer cell lines and FFPE derived-RNAof BC lesions was synthesized using BiozymcDNA synthesis kit. Variable concentrations (25ng, 50ng, and 100ng) of breast cancer cell lines and FFPE-derived total RNA were used for ESR1, PGR, and ERBB2 mRNA transcript detection using the corresponding primer pairs of each gene. GAPDH served as a reference gene. The following PCR program was applied: denaturation 950c (1min), 620c annealing temperature (20sec), 720c synthesis/extension temperature (20sec) for 35 cycles. The experiments were performed by including non-template controls in each PCR run. The PCR products were subjected to gel electrophoresis using SERVA agarose standard electro endosmosis (EEO)followed by documentation with the Image QuantTM LAS4000. Results: Multiple PCR experiments were carried out with different counts of PCR cycles, with and without extension, with various RNA concentrations and using breast cancer cell lines and FFPE-derived RNA. With the PCR conditions, testing of ESR1, PGR, and ERBB2 expression was possible. In particular, the MCF-7, MDA-MB-231 cell lines PCR run produced concordant data with IHC. However, some BC cell lines (BT-20 and SKBR-3) known as negative for the respective hormone receptor showed discordant results. Conclusion: In summary, the determination of the expression of ESR1, PGR and ERBB2 level in BC was successful. However, it requires further tests using different PCR conditions and a large set of known human breast tissues. From the experiment, we have learnt that endpoint PCR based ESR1, PGR, and ERBB2 detection can be used as an alternative method for better BC receptor diagnosis, treatment plan, and management in resource-limited countries. Citation Format: Zelalem Desalegn Woldesonbet, Martina Vetter, Meron Yohannes Nigussie, Tamrat Abebe Zeleke, Yonas Bekuretsion, Mahlet Arayselassie, Mathewos Assefa, Abebe Bekele, Endale Anberber1, Claudia Wickenhauser, Eva J Kantelhardt, Jürgen Bukur, Seliger Barbara. Optimization of end point PCR for the determination of ESR1, PGR, and ERBB2 expression level in resource-limited settings: Ethiopian context [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr A118.

Introduction: The burden of breast cancer is escalating across the globe and becoming a major public health problem in Ethiopia. Considering the fact that breast cancer is a heterogeneous disease in its nature, it demands a coordinated multimeric approach for diagnosis, treatment and management. Therefore, NanoString nCounter-based gene expression analysis might be employed for breast cancer intrinsic molecular subtyping, since it is a robust, highly reproducible tool and allows to simultaneously explore the expression of hundreds of genes in a single reaction tube. The aim of the study was to determine the intrinsic molecular subtypes of breast cancer samples from Ethiopian patients using a tailored NanoString nCounter gene expression assay for formalin-fixed, paraffin-embedded (FFPE) material. Methods: The present study was carried out to determine the molecular subtypes of breast cancer in Ethiopian patients. Archived FFPE tumor samples from public hospitals in Ethiopia were retrieved and blocks with clinical and pathologic data were used. From each sample total RNA was extracted by miRNeasy kit, then total RNA concentration and quality were determined and high-quality RNA was subjected to NanoString-based nCounter gene expression analysis using a tailored assay consisting of molecular and immunologic markers. For each gene, gene-fold change expression is given when compared to the reference cut-off value. The study focused on the analyses of estrogen receptor 1(ESR1), progestrone receptor (PGR), erythroblastic oncogene B (ERBB2) genes expression and the intrinsic molecular subtypes done using PAM 50 assay. Statistical analysis was carried out using SPSS software. Results: A total of 164 FFPE breast cancer blocks were analyzed for protein (ER, PR and Her2) expression status and mRNA expression levels. 86/164 (52.4%) breast cancer patients were tumor grade 3. More than 50% of tumor samples were ER and PR positive. In contrast to hormone receptors, only 20 (12.2%) breast tumor samples were found to be positive for HER-2. With regard to the mRNA expression levels of the respective genes, 106 (64.6%), 43 (26.2%) and 143 (87.2%) of breast cancer lesions analyzed showed an upregulation of ESR-1, PGR and ERBB2 genes, respectively. The samples with ER and PR positive results demonstrated by immunohistochemistry (IHC) showed nCounter levels from -60 to 124 (median=7) and -379 to 9 (median= -2), respectively. Samples with HER-2 positive results by IHC showed nCounter levels from -1 to 60 (median=17.5). Using PAM50 algorithm, we found 32 (19.5%) luminal A, 33 (20.1%) luminal B, 24 (14.6%) HER-2 enriched, 29 (17.7%) basal and 17 (10.4%) normal type breast cancer. Conclusion: According to the RNA expression assay, the majority of breast cancer subtypes were luminal A and luminal B and the lowest was normal type. In addition, the large majority of the cases analyzed were hormone receptor positive. Thus, hormonal therapy is of high importance for breast cancer care in Ethiopia. Citation Format: Zelalem Desalegn Woldesonbet, Martina Vetter, Meron Yohannes Nigussie, Tamrat Abebe Zeleke, Yonas Bekuretsion, Mahlet Arayselassie, Mathewos Assefa, Abebe Bekele, Endale Anberber, Claudia Wickenhauser, Eva J. Kantelhardt, Jürgen Bukur, Barbara Seliger. NanoString nCounter-based gene expression assay for evaluation of breast cancer molecular subtypes in Ethiopian patients [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C031.