The annual conference of National Cancer Institute - Cairo University "Bridging Gaps in Oncology"

P1 Downregulation of hypoxia-inducible factor 1 α expression inhibits growth and enhances IGF1R inhibitor OSI906 sensitivity in head and neck squamous cell carcinoma cells Ashraf Khalil, Mark Jameson Department of Clinical Biochemistry and Molecular Diagnostics, National Liver Institute, Menoufia University, Shebin ELkom, Egypt; Department of Otolaryngology – Head and Neck Surgery. University of Virginia, Charlottesville, Virginia, USA Correspondence: Ashraf Khalil (ashkalil2010@gmail.com) Journal of the Egyptian National Cancer Institute 2022, 34(Suppl 1):P1


Background
Hypoxia-inducible factor 1 α (HIF-1α) is a central regulator for cells to adapt to low cellular oxygen levels, is also often overexpressed and activated in many human cancers. HIF-1α mediates the primary transcriptional response of a wide range of genes in response to hypoxia. Insulin-like growth factor-1 receptor (IGF-1R) is a cell membrane receptor involved in cell proliferation is expressed in the head and squamous cell carcinoma.

Aim
To detect the effect of HIF-1α downregulation on the sensitivity of squamous cell carcinoma (SCC) to small molecule IGF1R inhibitor OSI-906. Methods A lentivirus-mediated doxycycline-inducible pTRIPZ short hairpin RNA micro (shRNAmir) plasmid targeting HIF-1α was transfected into two head and neck squamous cell carcinoma (HNSCC) cell lines to silence HIF-1α expression and to assess the effect of its downregulation on cell proliferation and sensitivity to IGF1R inhibitor. A nude mice animal model was developed by the transfection of a luciferase gene into the inducible shRNA HIF-1α cells and growing tumors by direct inoculation of the cells in the tongue. Tumor growth was determined by detecting the bioluminescence signal by the advanced IVIS 200 technology (Figure 2d). Result Downregulation of the expression of HIF-1α (Figure1 a-b) led to an increase in the sensitivity of head and neck squamous cell carcinoma to the OSI-906 IGF1R small molecule inhibitor (Figure 2a-b) by a mechanism involving the increase in apoptosis (Figure 2c).

Conclusion
The preliminary findings suggest that co-targeting of IGF-1R and HIF-1α may represent a novel approach for resistant head and neck tumors.
This study received supported from the National Institutes of Health To study the frequency of C-MYC/BCL-2 in adult Diffuse Large B Cell Lymphoma (DLBCL) patients presented to Egyptian National Cancer Institute and Tanta Cancer Center and to classify these patients into GC & ABC according to CD-10, BCL-6 and MUM-1. Also to assess the correlation between the presence of these prognostic markers and treatment outcome, overall survival and disease free survival. Background Non-Hodgkin lymphoma is the 7th most commonly diagnosed cancer among men and women; diffuse large B-cell lymphoma represents 32% of non-Hodgkin's lymphoma (NHL) cases. In Egypt, NHL is the 5th most common cancer. Methods A prospective study that included 100 adult patients with newly diagnosed and pathological confirmed Diffuse Large-B Cell Lymphoma (CD20+) who were treated by R-CHOP regimen presenting to Egyptian National Cancer Institute and Tanta Cancer Center between the period of January 2014 and December 2016 and followed up till January 2019. Detection of CD10, BCL6, MUM1, C-MYC and BCL2 by immunohistochemistry as prognostic markers. The following data were studied: patient's characteristics, treatment outcome and prospective overall survival (OS) and disease free survival (DFS). The current study was approved by NCI Institutional Review board (IRB no. IRV00004025). An informed consent was obtained from each patient.

Results
Age ranged from 25-68 years (median age was 49 years). Male cases were 52(52%) while female were 48(48%). Most of our patients presented with advanced stage (42% stage III and 25% stage IV) and (45%) of patients had bulky disease. Extra nodal sites were detected in 42% and patients presented with B-symptoms were 59%. The revised IPI showed that 4% had low IPI, 63% had intermediate IPI and 33% had high IPI. Chemotherapy response showed that 78 patients (78%) had CR response. CR response was lower in the double hit group (45%) while there is no difference between germinal center and non-germinal center groups. Only 21 patients were relapsed. The most chemotherapy toxicity was neutropenia in 29 patients (60.41%) and vomiting in 19 patients (59.37%), most of them were grade I and II. The median disease-free survival (DFS) was 49 months while 2-year and 4-year DFS was 89.2 %, and 59.9 respectively. The median overall survival (OS) was 51.8 months while the 2-year and 4year OS was 86% and 85% respectively. The germinal center was 35 cases (35%) and activated B-Cell was 65 cases (65%). Patients with double hit were 20 cases (20%), only 5 patients of them were germinal center while 15 patients were activated B-cell. There were better DFS in germinal center group and in non-double hit group. Within the double hit patients there were no statistically significant differences in overall survival between both groups (germinal and nongerminal). There were better overall survival in non-double hit group. Independent factors affecting response to chemotherapy were bone marrow involvement, extranodal involvement, IPI and bulky disease while those affecting disease free survival were germinal center and non-double hit groups and independent factors affecting overall survival were extranodal involvement and IPI.

Conclusion
Patients with double hit lymphoma had poor overall survival and disease free survival. There is no difference in overall survival between germinal center and non-germinal center group while the disease free survival was better in germinal center group. The CR response was lower in double hit group while there is no difference between germinal center and non-germinal center group. Keywords Diffuse large B-cell lymphoma, double hit lymphoma, germinal center and Activated B-cell Ethical committee approval: The current study was approved by National Cancer Institute Institutional Review board (IRB no. IRV00004025). Consent to participate: An informed consent was obtained from each patient.

Figure 2
Downregulation of HIF-1α suppress cell growth, colony formation, increased apoptosis, and the sensitivity of cells to IGF1R inhibition. A. Cells transfected with the silencing shRNAmir HIF-1α were treated with 2 μg/ml doxycycline for three days, then with OSI901 as indicated, and cell proliferation was determined at 72 h using alamarBlue. B. Colony formation assay at 14 days. C. Apoptosis assay 72h by Annexin V and PI by flow cytometry. D. Tumor detection and quantification by Bioluminescence imaging. The overall survival for adolescents and young adults (AYA) with Acute Lymphoblastic Leukemia (ALL) continues to lag behind that of younger children. Many retrospective and prospective studies reported better outcomes with the pediatric-inspired protocols when compared to the adult protocol. Aim This retrospective study aims to evaluate the efficacy and safety of adopting the pediatric-inspired ALL chemotherapy protocol in treating AYA patients diagnosed with ALL and to compare the clinical outcome with other adult ALL chemotherapy Protocol.

Results
Through this period, 169 patients were assessed and treated. The median age was 26years (range 18-39 years) with male to female ratio of 1.86:1. Most of the patients (n=108) had received the modified Dana Faber Protocol. Seven patients had received the Total XV protocol, while 54 cases had received modified GMALL protocol. B-cell ALL was reported in 120 patients (72.7%), of which(C-ALL: 18.8%, Pre B: 50.3%, Pro B: 3.5%), while 45 patients (27.3%) diagnosed with T-cell ALL (Early T: 14.5%, Intermediate T: 10.9% and Late T: 1.8%). Molecular and cytogenetic analysis for patients with B-ALL showed t(9;22) in 26.7%, t (4;11) in 4.2% while t (12;21) in one case only(0.8%). The pediatric-inspired regimens were well tolerated among our patients. When compared to the adult-protocol, the pediatric-inspired regimens were associated with significantly higher DFS (33 vs 18.8 months, p=0.046), While the OS was (26 vs 21 months, p=0.176), and CR rates (91% vs 86%, p=0.515). Also, the pediatric-inspired regimens were associated with lower incidence of septic shock during induction (24% vs 39%, p=0.013), lower incidence of early mortality (20% vs 33%, p=0.059), and shorter period of hospital stay during induction (Mean 31 vs 40 days, p<0.001). Conclusion -The pediatric-inspired chemotherapy regimens were well tolerated for AYA patients, and it was associated with better outcomes. -AYA patients should be treated as a unique category of ALL patients with major concerns regarding treatment-related longterm adverse events. -Continuous sincere efforts should be paid towards improving patients' adherence to treatment and management of treatment-related adverse events.

Keywords
Leukemia, Lymphoblastic, Adolescents and Young Adult Ethical committee approval: The study was approved by the Institutional Review Board (IRB) NCI-Cairo University. Consent to participate: it is a retrospective study with no intervention needed, no special informed consent but the study approval consent was taken by NCI Cairo University.

Results
According to MNA score: 43.7% of all patients were malnourished; 49.5% of stomach and 39.2% of colorectal cancer patients. Colorectal cancer cases had better survival rates than stomach cancer cases (p< 0.001). The independent factors that significantly affect OS were the cancer type and nutritional status. Tumor type was the only independent factor affecting PFS.

Conclusion
Malnutrition is higher in stomach cancer cases than in colorectal cancer cases, but the difference was not statistically significant. Colorectal cancer cases had better overall survival rates than stomach cancer cases. Malnourished had a worse survival than well-nourished. Keywords Malnutrition; Mini Nutritional Assessment; Gastro-Intestinal Cancer, Survival Ethical committee approval: The study was approved by the Institutional Review Board at NCI, CU (approval No: 201617063.3). Consent to participate: All participants signed the informed consent.

Background
The infectivity of COVID-19 and the variation of clinical behavior including fatal cytokine storms are considered one of the challenges nowadays. Recent reports showed that patients with an active or ongoing treatment for cancer are at a high risk of infection and COVID-19-related complications. Methods 49 patients admitted to NCI with different tumor types infected with COVID-19 were investigated as regards the clinical, pathological and laboratory characteristics, associated risk factors, as well as the course of hospital admissions (Tables 1-2 ).

Results
Patients with hematological malignancies represented 30/49 (61.2%) ( Table-1). The current study showed that 16 (32.7%) had multisystem organ failure and septic shock. Mortality was encountered in 14 patients (28.6%) within 15 (IQR: 2-34) days of covid-19 infection. Patients who had median serum ferritin of 2950 (range1000-10000) ng/L, showed increased incidence of mechanical ventilation and death. Similarly, patients with increased D dimer to a level of 1.2 (.9-1.3) ng/ml, had a significantly increased incidence of death. Patients with progressive or active cancer stages showed almost the same rate of COVID-19 infection in comparison to patients with stationary, remission or regressive disease. ICU admission was significantly increased in cancer patients infected with COVID-19 after surgery or who were on chemotherapy ( Table-3).

Conclusions
We can conclude that cancer patients infected with COVID-19 are at increased risk of adverse events and mortality, especially those receiving chemotherapy or underwent surgical procedure. Therefore, COVID-19 cancer patients should attain special care regarding the anticancer treatment and the other supportive measures to augment their immune system. Initiation of proper cancer treatment modality in newly infected patients with COVID-19 should mainly depend on the severity of viral infection, clinical condition, and finally oncological emergencies. Deaths in cancer patients with concomitant COVID-19 infections was mainly attributed to associated intensified treatment as well as gram negative sepsis. Keyword Cancer, mortality, COVID-19 Consent: Informed consent to participate in this study was obtained from participants (or their parent or legal guardian in the case of children under 16). IRBethical committee approval: Project funded by Cairo University number :10/2020 -Ethical committee approval at the National Cancer Institute, Cairo University: May/2020. Conflicts of interest: I have no potential conflict of interest to report-     The internalization of agonist activated G-protein coupled receptor (GPCRs) is a part of receptor trafficking. Also, it involves a list of endogenous proteins, where the endosomed ligand-receptors are distant to recycling or degradation as a mechanism of cellular signal management. Dynamins (Dyns) are large GTPase involved in multiple endocytic pathways, including receptor mediated endocytosis. They perform unique tasks including endosome fission, mitochondrial severing and some signaling events. The impact of Dyns inhibition in vasopressin stimulated or AKT/mTOR inhibited breast cancer cells was not addressed. To explore this, Dyns were selectively inhibited by dynsore in triple negative breast cancer cells (MDA MB-231) in which vasopressin receptor-2 (VR2) was agonist activated with exogenous arginine vasopressin hormone (AVP) or PI3K/Akt/mTOR was inhibited by Wortamnin (Wort). The results depicted the cytotoxic effect of Dyn inhibition, where Dynasore, individually promoted autophagic and apoptotic cell death. Apoptosis was developed in 19%, whereas the autophagy marker LC3II protein was observed in 39%. Dyns inhibition in AVP stimulated cells, PI3K/mTOR inhibited cells or both progressively enhanced the apoptotic and autophagic effects. Similarly, 73% of cells were arrested in G0/G1 phase. The activation of Akt into P-Akt decreased by Dynasore and more repression was observed in cells dually treated with Dynasore, AVP, Wort or both. Also, the drug was able to minimize the invasion of cells as indicated by the transwell assay.
Conclusively the results presented Dynamins as anticancer targets in invasive breast cancer cells.

Background
The outcome of childhood Burkitt's lymphoma (BL) has improved steadily over the past decades through the use of intensive sequential multi-agent chemotherapy regimens with increased concern about increasing toxicity related mortality.

Aims
This study objective was to assess incidence and risk factors of early mortality in BL   is of huge concern. It takes months and even years for getting a potential anti-viral agents.
In this work, we develop an effective confirmatory immunoassay based on using monoclonal antibody and a secondary labelled antibody, to evaluate the anti-viral activity of any suggested antiviral agent against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), avoiding laborious culture technique which is not available in many Labs in Egypt. Chitosan nanoparticles (CNPs) were selected to evaluate our newly immunoassay after evaluating its antiviral effect in both in-silico and in-vitro studies.

Methods
Docking of CNPs was carried out against spike protein of SARS-CoV-2 and binding scores were calculated. CNPs were prepared using ionic gelation method. They were characterized using Scanning Transmission Electron Microscopy (STEM), Fourier-transform infrared spectroscopy (FT-IR), and Zeta analyzer. Cytotoxicity was performed on Vero E6 using MTT colorimetric assay. Antiviral activity was evaluated using real-time polymerase chain reaction (PCR) assay. The binding affinity between the Spike protein and CNPs was evaluated and assessed using a monoclonal antibody and a secondary antibody conjugated to Alexa Flour 647 dyes protocol. The viral inhibition was assessed using varying concentrations of CNPs (1, 5, 10, 20, 30, and 50 μg/ml).

Results
The strength of binding interactions between spike protein and CNPs ligand complexes gave scores at -6.6 Kcal/mol which is the same for Remdesivir (reference drug). CNPs were prepared at sizes of 35 nm with surface charges of 42 mv. The cytotoxic concentration of CNPs that cause death to 50 % of viable cells (CC50) was determined at 135 μg/ml on Vero E6, cellular uptake confirm localization of nanoparticles inside cells. Antiviral activity of CNPs against SARS-CoV-2 reached at 10 μg/ml as evidenced by undetected viral copies/ml using quantitative real-time PCR assay.

Conclusion
These results demonstrate that CNPs represent a promising antiviral candidate against entry of SARS-CoV-2 and successfully preventing infection.
-Funding Agency: ASRT-project ID 6901 -There are no clinical samples involved in this research, so the research did not require an approval from ethics committee.

P12
Predictors and outcome of infection related mortality in pediatric acute myeloid leukemia, febrile neutropenic episodes analysis, single institute experience  Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
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