Specific targeting of type I interferon activity to the tumor microenvironment or to dendritic cells as a novel, generic and safe cancer immunotherapy Anje Cauwels1, Sandra Van Lint1, Alexander Van Parys1, Franciane Paul2, Geneviève Garcin2, Stefaan De Koker1,5, Sarah Gerlo1, Yann Bordat2, Bart Vandekerckhove4, Gilles Uzé2 & Jan Tavernier1,3 1Cytokine Receptor Laboratory, VIB Medical Biotechnology Center & Ghent University, Gent, Belgium. 2CNRS UMR 5235, University Montpellier, Montpellier, France. 3Orionis Biosciences, Zwijnaarde-Gent, Belgium; and Watertown, Boston, USA. 4Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, UZ Gent 5Present address: eTheRNA Immunotherapies, Belgium Since a couple of years, immunotherapy has been set forth as a fourth pillar for cancer treatment, next to surgery, chemotherapy and radiotherapy. Immunotherapeutics include checkpoint inhibition targeting antibodies, several T lymphocyte and Dendritic Cell (DC) based cellular therapies, and cytokines. Type I IFN has been approved for the treatment of several solid and blood cancers. However, best results are usually obtained with higher doses, and the many systemic toxic side effects of IFN thus are severely dose-limiting. To curtail cytokine toxicity, we developed AcTakines, Activated-by-Targeting Cytokines, improved (mutated, with reduced receptor affinity) immunocytokines fused to cell-specific targeting moieties. As a murine AcTaferon (type I IFN based AcTakine), we use hIFN2-Q124R, which breaches the strict cross-species barrier and is thus weakly active on mouse cells. Selective targeting is achieved by coupling the AcTaferon to single domain antibodies (sdAbs) recognizing cell-specific surface markers. Targeting AcTaferon with CD20 sdAb to CD20+ cells, such as B lymphocytes or A20 B cell lymphoma, restored full activity in vitro as well as in vivo. In vivo treatment of A20 or B16-CD20+ tumors with CD20-targeted AcTaferon drastically reduced tumor growth, similar to high dose wild-type mIFN immunocytokine. In sharp contrast to the latter, however, it did not cause any systemic toxicity (evaluated via body weight, temperature, and blood cell counts). The AcTaferon antitumor effect was lost in IFNAR1-deficient, Batf3-deficient or CD8-depleted animals, as well as in mice lacking IFNAR1 on CD11c+ cells, indicating involvement of conventional DC (cDC). Furthermore, selective targeting of AcTaferon to Clec9A+ XCR1+ cDC1 was sufficient to induce tumor stasis in both mouse melanoma and breast carcinoma models, as well as in a human lymphoma model in humanized mice. When combined with immunogenic chemotherapy, low-dose TNF, or immune checkpoint blockade, complete tumor regressions and long-lasting tumor immunity could be observed, still without adverse effects. Collectively, our findings thus indicate that tumor- or DC-targeted AcTaferons provide novel, highly efficient and safe cancer (immuno)therapies, synergizing with existing therapies to completely eradicate tumors, and capable of converting immunogenically “cold” tumors into “hot” tumors. In case of DC-targeting, there is no need for tumor-specific markers or ex vivo cell manipulations, in contrast to other currently used cellular therapies, and generic application is possible in a broad range of malignancies.
Prof. Jan Tavernier obtained his Ph.D. degree in 1984 on the cloning of interferon and interleukin genes. After an extended stay at Biogen and later at Roche, he returned in 1996 to Ghent University at the VIB Department of Medical Protein Research. He founded the Cytokine Receptor Laboratory that currently hosts over 30 researchers. His main areas of expertise are cytokine receptor activation and signal transduction, and the analysis of protein-protein interactions including interactome mapping, pathway walking and molecular description of inter-domain interactions (www.mappit.be). More recently, he is also developing the AcTakine concept: immunocytokines which are devoid of any systemic toxicity.Jan Tavernier published more than 250 refereed manuscripts, 20 of which are being cited over 100 times. He also holds 35 patent applications. He is Chair of the Department of Biochemistry at the Faculty of Medicine and Health Sciences and is Co-Director of the VIB Department of Medical Protein Research. He is a member of Royal Belgian Academy of Sciences and The Arts.
Fundamentals in Translational oncology research and its clinical importance Presentation: Oral Name: motawa E. El-Houseini Adress: Dept. Cancer Biology, NCI.Cairo University, Egypt Mail id: email@example.com Pn No: 01112139714 Abstract Translational oncology research( TOR ) is a cross- talk between two players: A- Lab.-based researchers (scientists) They do in-vitro experiments in test tubes , culture flasks or Petri dishes and in-vivo experiments using animal models B- Clinicians: They do clinical trials where patients are directly involved in the testing of a substance or treatment that the formers have introduced. TOR aims to: -Bring the two dimensions together -(Lab.-based researchers and clinicians ) -Make the lab.work relevant to clinical practice so as to introduce novel therapy or biomarkers for an early detection, diagnosis and prognosis for malignancies
Motawa E. El-Houseini is currently working as Professor of Biochemistry and Molecular Biology - Cancer Biology Department, Cairo University, Egypt. His Research on Evaluation of basic fibroblast growth factor and matrix metalloproteinases in breast and bladder cancer patients. Professor and Chairman of Cancer Biology Dept .(1998 - 2001),He was Research Fellowship from DANIDA at Fibiger Laboratories, Finisin Institute, Copenhagen, Denmark (1979 – 1983), And he was a NIH Research Fellowship at Pittsburgh cancer institute, Transplantation institute
Cancer: An Ancient Disease and Modern Treatment; Utmost necessity for a nontoxic anticancer drug Prof. Manju Ray# Honorary Visiting Professor Department of Biophysics, Bose institute, Centenary Building P-1/12, CIT Scheme VII-M, Kolkata- 700 054, India Telephone no. - +919830108934(M); 0913323553886 (fax). Email address- firstname.lastname@example.org; email@example.com # Distinguished Professor GLA University, Mathura, India Cancer is an ancient deadly disease causing worldwide mortality. Most chemotherapeutic agents follow non-specific distribution and are associated with severe side-effects. Our concern was to develop an anticancer therapeutic(s) that kill(s) the cancer cells without affecting healthy normal cells. Methylglyoxal, an enigmatic ketoaldehyde formed as a glycolytic intermediate has been well studied for its potent anti cancer activities. We deciphered through a series of investigative experiments that MG mediates anti cancer activities by inhibiting glycolysis and mitochondrial respiration of exclusively malignant cells. Detailed pharmacokinetic and toxicological studies on MG confirmed with no toxicity to normal cells. We obtained due permission from the regulatory authorities of India to carry out Phase I/II trial over a period of 10 years in three divided study groups with 105 terminally ill cancer patients. Among the 105 patients treated with methylglyoxal, complete remission was observed for 48 patients and partial remission for 26 patients. Among the remaining patients at least 12 patients lost follow-up. The treatment was found to be especially effective for adenocarcinoma of urinary bladder, breast, uterus, esophageal and gastrointestinal tract cancer. Several vital biochemicals, radiological and other parameters were tested in patients who received treatment for a long time implicated no toxicity as per the parameters studied. Most importantly, it ceases bleeding and relieves pain. Now the drug is approved for Phase III trial by regulatory authorities and waiting for funding. Modern day chemotherapy is taxed with adverse side effects and often leads to life threatening complication. The advantage with MG is its ability to target and kill neoplastic cells without affecting normal cells at the therapeutic dose and shows great promise in the development of anti cancer based therapeutics.
Manju Ray is an Indian scientist in Molecular Enzymology and Cancer Biochemistry. She has done notable work in the development of anticancer drug and understanding of differentiation process of cells. Her interests cover tumor biochemistry and molecular enzymology.She was awarded the most prestigious Shanti Swarup Bhatnagar Prize in Biological Sciences for the year 1989. She also got Young Scientist Medal, Life Time Achievement Award, Dr I.C. Chopra Memorial Award, Dr. Jnan Chandra Ghosh Memorial Award etc. Prof Ray has 4 patents (US and PCT).She delivered many lectures in several Research Institutes and Universities in India and abroad like USA, England, Sweden, Australia, China, Germany, Canada etc. She has published total 76 papers in international reputed journals.
Feijoa sellowiana fruit, an amazing source for anti-cancer molecules Adriana Basile1 1 Dipartimento Biologia, University of Naples Federico II, Napoli, Italy Feijoa sellowiana (F. sellowiana or Acca sellowiana), also known as pineapple guava or guavasteen, is a subtropical species of the Myrtaceae family. Different biological activities have been reported for this fruit and its components (Basile et al., 1997; 2010). Topic of this report is the anticancer activity tested on a variety of cellular models. The Feijoa acetonic extract exerts anti-cancer activities on solid and haematological cancer cells. The extract did not show toxic effects on normal myeloid progenitors, thus displaying tumour-selective activity. In the Feijoa acetonic extract, fractionation and subsequent purification and analyses identified Flavone as the active component. Flavone induces apoptosis, which is accompanied by caspase activation and p16, p21 and TRAIL overexpression in human myeloid leukaemia cells. Use of ex vivo myeloid leukaemia patient blasts confirms that both the full acetonic Feijoa extract and its derived Flavone are able to induce apoptosis. In both cell lines and myeloid leukaemia patient blasts the apoptotic activity of Feijoa extract and Flavone is accompanied by an increase in histone and non-histone acetylation and by HDAC inhibition. The Feijoa apoptotic active principle is Flavone, whose activity correlates with the induction of HDAC inhibition, supporting the hypothesis of its epigenetic pro-apoptotic regulation in cancer systems. Finally we are investigating the effects of F. sellowiana extract on mesenchymal stem cells from adipose tissue. In particular we are evaluating differentiation of mediastinal cells characterizing adipogenesis, chondrogenesis, ossification by an approach based on differentiation test and FACS. Biography Adriana Basile is Associate Professor in Botany at the Faculty of Mathematical Physical and Natural Sciences, University of Naples, Federico II. Member of the Faculty Council for the Interdepartmental PhD in Monitoring and mitigation of environmental risk. She was the director of CISME, Interdipartimental Cemnter of Electron Microscopy, Member of Italian Botanical Society and American Botanical Society, and International Association of Bryologists. Guest editor of Molecules and peer reviewer of Natural Product Research, Pharmaceutical Biology, International Journal of Molecular Science, Journal of Medicinal Plant Research, Food and Chemistry Toxicology, Journal of Pharmacy and Pharmacology, Jornal of Plant Physiology, Chemoterapy.
Adriana Basile is Associate Professor in Botany at the Faculty of Mathematical Physical and Natural Sciences, University of Naples, Federico II. Member of the Faculty Council for the Interdepartmental PhD in Monitoring and mitigation of environmental risk. She was the director of CISME, Interdipartimental Cemnter of Electron Microscopy, Member of Italian Botanical Society and American Botanical Society, and International Association of Bryologists. Guest editor of Molecules and peer reviewer of Natural Product Research, Pharmaceutical Biology, International Journal of Molecular Science, Journal of Medicinal Plant Research, Food and Chemistry Toxicology, Journal of Pharmacy and Pharmacology, Jornal of Plant Physiology, Chemoterapy
Title:Development and control of mouse lung tumors in the offspring from the predisposed mothers. Presentation: Oral Name:Krishna P. Gupta Address:B.B.A. University,Lucknow. Mail id: firstname.lastname@example.org Pn No:09838106618 Epidemiological and experimental studies suggest that the exposure to environmental carcinogens during the pregnancy increased the susceptibility to develop cancers in the offspring. Risk of cancer development in the offspring due to exposure of carcinogen during pregnancy is alarming and needs to be addressed. N-ethyl-N-nitrosourea (ENU) induced mouse lung tumor model showing histological and molecular similarities to human lung adenocarcinoma, has been used to study the transplacental tumorigenesis in the mouse lungs. The status of PI3K/PTEN/Akt pathway and microRNAs in F1 mice from the ENU exposed mothers during the gestation period and their response to inositol hexsphosphate (IP6)was looked upon. IP6 prevented the alterations observed in F1 mice from the ENU exposed mothers. We show that the alterations in PI3K/PTEN/Akt pathway along with associated micro RNAs are clearly involved in ENU caused transplacental lung tumor development in F1 mice. Suppression of this altered pathway by IP6 could be reducing the lung tumor development and thus, could be exploited further as potential targets for cancer control. We are providing a rationale targeting multiple pathways in the management of tumors in general and lung tumors in particular in second generation. Biography Dr. Krishna P. Gupta did her PhD at the King George’s Medical College, Lucknow, India. She was a postdoctoral fellow and also visiting scientist at Baylor College of Medicine, Houston; M.D. Anderson Cancer Centre, Houston , Institute of Toxicology, University of Mainz, Germany; Buffalo State and Roswell Park Cancer Institute, Buffalo, NY, USA. She worked as scientist and the professor in carcinogenesis, CSIR-Indian Institute of Toxicology Research, Lucknow for more than thirty years. She has published in peer reviewed international journals of repute , has US patent and is in the editorial boards of the scientific journals. Currently Dr. Gupta is working as Emeritus Medical Scientist at B.B.A.University, Lucknow,India.
Dr. Krishna P. Gupta did her PhD at the King George’s Medical College, Lucknow, India. She was a postdoctoral fellow and also visiting scientist at Baylor College of Medicine, Houston; M.D. Anderson Cancer Centre, Houston , Institute of Toxicology, University of Mainz, Germany; Buffalo State and Roswell Park Cancer Institute, Buffalo, NY, USA. She worked as scientist and the professor in carcinogenesis, CSIR-Indian Institute of Toxicology Research, Lucknow for more than thirty years. She has published in peer reviewed international journals of repute , has US patent and is in the editorial boards of the scientific journals. Currently Dr. Gupta is working as Emeritus Medical Scientist at B.B.A.University, Lucknow,India.
Emerging Trends in Epidemiology of Breast Cancer: Current Scenario and Challenges Girish Sharma1, 2 1Amity Center for Cancer Epidemiology & Cancer Research, 2Amity Institute of Biotechnology, Sector 125, Amity University Uttar Pradesh, NOIDA, India Email: email@example.com; firstname.lastname@example.org Breast cancer is the most frequently diagnosed malignancy in women globally. Its incidences have increased by more than 20%, while mortality rate has increased by 14%. The increasing trends of breast cancer, in both developed and developing countries, are an actual threat. In 2012, globally around 1.67 million breast cancer cases were diagnosed (25% of all cancers), and surprisingly 5.2 million cases of mortality were recorded, thereby making it the leading cause of death among women. Incidence rates in Asian women have been rising which is mainly linked with a shift towards a more westernized lifestyle. In India also, the incidence of breast cancer is rising. It is estimated that in 2008 there were 115,251 new cases of breast cancer with an age standardised incidence rate (ASR) of 22.9 per 100,000. Furthermore, by the year 2030 the number of new cases of breast cancer in India is estimated to reach approximately 200,000 per year. Variation of rate of incidences is almost four-fold across the world regions, the difference in the rates ranges from 27 per 100,000 in Eastern Asia and Middle Africa and 96 per 100,000 in Western Europe. The mortality rate is low in world regions because of the more favourable survival rate of breast cancer has been noted in developed regions, with rates ranging from 6 per 100,000 in Eastern Asia to 20 per 100,000 in Western Africa. Higher consumption of fats and reduced or no consumption of fruits and vegetables along with smoking, consumption of alcohol has been shown to increase the risk of breast cancer. Sedentary lifestyle and lack of exercise also add to the risk. Age, Race/Ethnicity and family history are the most well established risk factors associated with development of breast cancer. Mutations in the BRCA1 and BRCA2 genes have been identified to be strongly associated with the risk of developing breast cancer. Stress induced aberrations resulting in lifestyle changes amongst women of the Indian sub-continent along with enhanced genetic predisposition is probably one of the predominant reasons for higher rates of breast cancer. The peak age of breast cancer occurrence has shifted towards younger women. The presentation shall discuss current scenario and challenges in the management of breast cancer. Keywords: Breast cancer, incidence, mortality, risk factors.
A Novel Potential Effective Strategy for Enhanced Antitumor Immune Response in Breast Cancer patients Using Viable Cancer Cell-Dendritic Cells Based Vaccine Mona S. Abdel Lateif1, Sabry M.Shaarawy1, Eman Z. Kandeel2,Ahmed H.El-Habashy3, Mohamed L. Salem4, Motawa E. El-Houseini1 1Cancer Biology Department, National Cancer Institute, Cairo University 2Clinical Pathology Department, National Cancer Institute, Cairo University 3Pathology Department, National Cancer Institute, Cairo University, 4Zoology Department, Faculty of science, Tanta University. Abstract Dendritic cells (DCs) have been used in many clinical trials for cancer immunotherapy, but they have met with a limited success in solid tumors. So the aim of this study was searching for a novel potential effective immunotherapy for breast cancer patients through in vitro optimizing conditions of viable cancer cell - dendritic Cells based Vaccine. Immature DCs were primed by viable MCF-7 breast cancer cells. We assessed; DCs activity and maturation through measuring CD83+, CD86+ and MHC-II, as well as, different T cell subpopulations namely; CD4+,CD8+ and regulatory T-cells CD4+CD25+ FOXP3+ by flowcytometric analysis. Moreover, we measured Fork head box protein 3(FOXP3+) level by ELISA, as well as FOXP3 gene expression by real time PCR. The levels of interleukin 12 (IL12) and interferon gamma (IFNγ) were determined by ELISA. Finally, we evaluated the cytotoxicity of cytotoxic T cells (CTLs) through measuring Lactate dehydrogenase (LDH) release assay by ELISA. Our results revealed that, CD83+, CD86+ and MHC-II on DCs, were significantly elevated (P value <0.001) after priming with breast cancer cells. In addition, there was an activation of T-cells CD4+ and CD8+, with a significant decrease of CD4+CD25+ FOXP3+T- regulatory cells (P value <0.001). Furthermore, the latter did show a significant down-regulation of FOXP3 gene expressions (P value <0.001) and a significant decrease in the level of its protein after activation (P value <0.001). Meanwhile, a significant increase in the secretion of both IL12 and IFN-γ (P value= 0.001) was observed. LDH release was significantly increased (P value <0.001) indicating a marked cytotoxicity of CTL against cancer cells. Therefore viable breast cancer cell - DCs based vaccine could open new avenue for a novel breast cancer immunotherapy.
Abstract The present review systematically investigated how natural taurine exerts a dual role to regulate either apoptotic or anti-apoptotic biological networks as a protective mechanism. This review clarify that taurine protect different type of tissue through various pathways, differs according to each tissue disordered and its underline causes. Taurine was reported to possess anti-apoptotic effect against oxidative stress-induced apoptosis in several cells, such as hepatocytes cardiomyocytes and epithelial cells. On the other hand it exerts an apoptotic effect concerning numerous cancerous cells and other dangerous cells such as HSCs protecting the liver from hepatic fibrosis. Taurine was also found to synergise and potentiate the anti-proliferative effects of many chemotherapeutic drugs, by enhancement of tumor cell apoptosis selectively, while acting by its immunomodulation effect to attenuate lymphopinia by its anti-apoptotic effect. Throughout this review we found that taurine regulate apoptosis mechanism depending on which was beneficial to the organisms through the regulation of different apoptotic signaling pathways as well as direct effects on the regulation of pro-apoptotic proteins at thier expression level. This review could help casting a new light on the action mechanism and targets of taurine as a natural protective agent. Due to the nature of taurin as an endogenous anti-oxidant synthesized by the human body we can consider taurine as promising and safely agent that might use to prevent and attenuate the adverse effects of many diseases, especially cancer that can be used as a prophylactic or therapeutic agent. Biography Dr. Omhashem M. abd-elhameed has completed her PhD on Cancer biology and biochemistry from Cancer Biology Department, Biochemistry and Molecular Biology Unit, National Cancer Institute, Cairo University-Egypt, under supervision of Proff. Dr. Motawa E. EL-Houseini. Postdoctoral studies continued with Dr Motawa E. EL-Hous ini and Dr wafaa Ahmed ; on the same institute. I work as lecturer on biomedical engineering department of the higher institute of engineering- elshrouk- Cairo-Egypt. , and member of internal audit committee in the quality unit of the Institute of Engineering. I have published about 5 papers in reputed journals and now we prepare for a project ((phase 2 clinical trial for treatment of HCC, using some natural products. this work will be a chair between National cancer institute and National Hepatology and Tropical Medicine Research Institute in Egypt. My research program target two main points; The early detection of cancer and Access to the best use for natural materials that can be prevent and treat cancer development to avoid the negative symptoms resulting from the use of traditional methods of treatment. Also I am interested looking for the use of nanotechnology for improving the detection and treatment of cancer.
Dr. Omhashem M. abd-elhameed has completed her PhD on Cancer biology and biochemistry from Cancer Biology Department, Biochemistry and Molecular Biology Unit, National Cancer Institute, Cairo University-Egypt, under supervision of Proff. Dr. Motawa E. EL-Houseini. Postdoctoral studies continued with Dr Motawa E. EL-Hous ini and Dr wafaa Ahmed ; on the same institute. I work as lecturer on biomedical engineering department of the higher institute of engineering- elshrouk- Cairo-Egypt. , and member of internal audit committee in the quality unit of the Institute of Engineering. I have published about 5 papers in reputed journals and now we prepare for a project ((phase 2 clinical trial for treatment of HCC, using some natural products. this work will be a chair between National cancer institute and National Hepatology and Tropical Medicine Research Institute in Egypt. My research program target two main points; The early detection of cancer and Access to the best use for natural materials that can be prevent and treat cancer development to avoid the negative symptoms resulting from the use of traditional methods of treatment. Also I am interested looking for the use of nanotechnology for improving the detection and treatment of cancer.
Diagnostic and prognostic value of circulating microparticles in small cell lung cancer Fadi Najjar1, Moosheer Alammar2, Ghassan Al-Massarani1, Adnan Ikhtiar3 1Biomarkers Laboratory, Radiation Medicine Department, Atomic Energy Commission of Syria (AECS), Syria 2Division of Thoracic Oncology, Oncology Department, Albairouni University Hospital, Syria 3Radiobiology laboratory, Biotechnology Department, Atomic Energy Commission of Syria (AECS), Syria Abstract Circulating microparticles (MPs) have been proposed for the assessment of non-small cell lung cancer (NSCLC). However, little is known about their potential clinical relevance in small cell lung cancer (SCLC). The aim of this prospective study was to analyze whether circulating MPs could be used as complementary biomarkers in the clinical setting of SCLC. Forty-nine patients with SCLC and 41 healthy subjects were included in this prospective study. Thirty-seven patients presented at initial diagnosis (ID) and 12 patients at relapse (R). Measurement of circulating MPs was performed using flow cytometry technique. Pretreatment MPs levels were significantly higher in SCLC patients either at ID or R than those in healthy subjects (p < 0.007). Baseline serum MPs levels were not significantly correlated with clinicopathological parameters including age, BMI, estimated weight loss, smoking degree and staging (all p > 0.05). However, estimated tumor volume (ETV) was inversely correlated with basal MPs values (p < 0.0001). Moreover, patients with an objective response (OR) displayed significantly decreased pretreatment MPs levels (p = 0.05). Using receiver operating characteristics (ROC) curves the cut-off value of baseline MPs number was 1257 events/µL, with sensitivity and specificity of 71.4% and 83%. Pretreatment MPs values were significantly higher in one-year survivors than in one-year nonsurvivors (p = 0.05). Circulating levels of MPs are significantly increased in SCLC patients as compared with healthy subjects. Furthermore, baseline MPs number seem to be considered as new biomarker for predicting tumor response and long-term survival in SCLC. Keywords: Biomarkers, Circulating microparticles, lung cancer.
1β-2-Himachalen-6-ol: A Novel Anticancer Sesquiterpene Unique to the Lebanese Wild Carrot Robin I. Taleba, Mirvat El-Sibai,a Costantine Dahera and Mohamad Mrouehb aDepartment of Natural Sciences, Lebanese American University, Byblos1102 2801, Lebanon bSchool of Pharmacy, Lebanese American University, Byblos 1102 2801, Lebanon Daucus carota ssp. carota, also known as wild carrot, is a commonly used herb in Lebanese folk medicine to treat several ailments including cancer. Previous studies in our laboratories showed that the Daucus carota oil extract (DCOE) and subsequent fractions exhibit antioxidant, anti-inflammatory and anti-cancer activities. In this study, we report the isolation and identification of the major compound responsible for the anti-cancer activity of DCOE along with the mechanism of action involved. GC-MS and NMR spectroscopy revealed the identity of the major compound as 1β-2-himachalen-6-ol, a novel sesquiterpene unique to the Lebanese wild carrot. 1β-2-Himachalen-6-ol demonstrated potent anti-cancer activity against B16F-10, Caco-2, MB-MDA-231, A549 and SF-268 cancer cells (IC50 13-4 μg/mL; 58-18 μM), with SF-268 cells being the most sensitive. The sesquiterpene was shown to induce cell death through apoptosis (flow cytometry), decrease 2D cell motility (wound healing assay) and 3D invasion, as well as increase cell adhesion in SF-268 cells. Additionally, 1β-2-himachalen-6-ol showed very low toxicity in mice with an LD50 > 6000 mg/kg body weight. In conclusion, the present data demonstrate that 1β-2-Himachalen-6-ol is a potential multi-mechanistic chemotherapeutic drug with high potency and safety. Key words: Daucus carota; 1β-2-Himachalen-6-ol; cytotoxicity; cell invasion; wound healing; cancer Biography Dr. Taleb is an Associate Professor in Medicinal Chemistry at the Lebanese American University (LAU). He obtained his PhD in Medicinal Chemistry from UWS (Australia) and has since published over 24 articles in the fields of cancer research, oncology, drug discovery and inorganic chemistry. Dr. Taleb has practical experience in instrumental analysis including NMR, HPLC, GC, Elemental Analysis, Mass-Spec, IR, UV, Protein Synthesiser, LCMS/MS and Circular Dichroism (CD). Dr. Taleb is a Member of the Royal Australian Chemical Institute (MRACI) as well as the American Chemical Society (ACS). Dr. Taleb is a reviewer for Elsevier Publishers.
Dr. Taleb is an Associate Professor in Medicinal Chemistry at the Lebanese American University (LAU). He obtained his PhD in Medicinal Chemistry from UWS (Australia) and has since published over 24 articles in the fields of cancer research, oncology, drug discovery and inorganic chemistry. Dr. Taleb has practical experience in instrumental analysis including NMR, HPLC, GC, Elemental Analysis, Mass-Spec, IR, UV, Protein Synthesiser, LCMS/MS andCircular Dichroism (CD). Dr. Taleb is a Member of the Royal Australian Chemical Institute (MRACI) as well as the American Chemical Society (ACS). Dr. Taleb is a reviewer for Elsevier Publishers.
Methanolic Extract from Aerial Parts of Artemisia Annua L. Enhances Sensitivity of Pre-B Acute Lymphoblastic Leukemia Cell Lines to Vincristine Pargol Mashatia, Somayeh Esmaeilib, Nasrin Dehghan-Nayeria, Mina Darvishia, Ahmad Gharehbaghiana,c* aDepartment of Laboratory Hematology and Blood Bank, Shahid Beheshti University of Medical Sciences, Tehran, Iran bTradintional Medicine and Materia Medica Research Center, School of Traditional Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran c Department of Laboratory Hematology and Blood Bank, Shahid Beheshti University of Medical Sciences, School of Allied Medical Sciences, Tehran, Iran Purpose: Vincristine (VCR) is a widely used chemotherapy drug in the treatment of acute lymphoblastic leukemia (ALL). Clinical use of VCR is highly limited by its neurotoxicity and dose-limiting effects. Combination therapy with natural products can be an efficient therapeutic approach to reduce adverse effects of chemotherapy drugs. The aim of the present study is to investigate the synergistic effect of methanolic extract from aerial parts of Artemisia annua in combination with VCR on leukemic cell death in vitro Methods: MTT assay was carried out to determine the cytotoxicity activity of the extract and VCR. Combination Index was evaluated to assess the synergism/antagonism effect of two agents. Apoptosis was analyzed by flow cytometry and Caspase 3 activity assay. mRNA expressions of Caspase 3, Bax, Bcl-2 were quantified using RT-PCR. One-way ANOVA and post hoc Tukey multiple comparison tests were used for statistical analysis. Results: Our results showed that the combination of Artemisia annua and VCR caused time and dose-dependent decrease in cell viability and induced apoptosis in both cell lines. Lower doses of VCR and the extract displayed significant synergistic inhibition of Nalm-6 and Reh cell growth (P< 0.001). Interestingly, concurrent use of A. annua and VCR resulted in Caspase 3 and Bax gene upregulation and Bcl-2 gene downregulation (P< 0.05). Conclusion: From the results presented here, the extract of Artemisia annua substantially increased sensitivity of leukemic cells to VCR, suggesting that this combination could potentially decrease VCR drug side effects.
Innovation in Photodynamic Therapy of Tumors by Lasers: An Overview of Forty years Experience from Drug Discovery to Bench and Clinical Applications. Mohamed El-Far Mansoura University, EGYPT We will present our experience for four decades innovations of photodynamic therapy (PDT) of certain types of tumors. Presentation will cover the followings: (1) our discovery of abilities of some natural porphyrins to be used as tumor localizers and photosensitizers as well as their uses for the first time in this field towards diagnosis and treatment of tumors by laser-PDT. (2) Mechanism of action and our innovations to increase the efficacy of PDT of tumors. (3) Potential use of 5-Aminolevulinic acid (5-ALA) in a newly developed technique for treatment of bladder cancer for first time and innovations about distribution and selective in-vivo tumor localization of endogenous porphyrins induced and stimulated by 5-ALA. (4) Preparation of novel effective photosensitizers for PDT. (5) Novel safe approach to eliminate photo-toxicity side effect. (6) Our vast experience in experimental as well as clinical applications of PDT of tumors. Biography Professor Mohamed El-Far, worked in biochemistry field for 40 years, published over 90 peer-reviewed papers. He received Fulbright and British council fellowships several times as well as German DAAD grant to establish PDT Program at Munich, also received US-AID grant to establish PDT unit in Egypt. He is serving on the editorial boards and Hon. Editor to four international journals. He acts as UNESCO expert in science and technology. Dr. El-Far served as visiting professor to University of California as well as Utah laser center also Mayo clinic for several years. He also served as a visiting professor to Cardiff and Swansea Universities, UK. He is a member of International Photodynamic Association and Royal Society of Chemistry, UK. Selected and served as expert and consultant for biochemistry in the national committee of suprime council of Universities in Egypt, which is the highest nation honor. Recently selected as a member of higher Education and Research committee for Mansoura University. Selected by the International Biographical Centre, Cambridge, England to be among top 100 scientists 2012. Received honary doctorate of Letters from International Biographical Centre, Cambridge, England. Selected on the list of global speakers as a keynote speaker and honorable guest for several international conferences in cancer and diabetes as well as other recent biomedical applications as stem cells and nanotechnology.
Professor Mohamed El-Far, worked in biochemistry field for 40 years, published over 90 peer-reviewed papers. He received Fulbright and British council fellowships several times as well as German DAAD grant to establish PDT Program at Munich, also received US-AID grant to establish PDT unit in Egypt. He is serving on the editorial boards and Hon. Editor to four international journals. He acts as UNESCO expert in science and technology. Dr. El-Far served as visiting professor to University of California as well as Utah laser center also Mayo clinic for several years. He also served as a visiting professor to Cardiff and Swansea Universities, UK. He is a member of International Photodynamic Association and Royal Society of Chemistry, UK. Selected and served as expert and consultant for biochemistry in the national committee of suprime council of Universities in Egypt, which is the highest nation honor. Recently selected as a member of higher Education and Research committee for Mansoura University. Selected by the International Biographical Centre, Cambridge, England to be among top 100 scientists 2012. Received honary doctorate of Letters from International Biographical Centre, Cambridge, England. Selected on the list of global speakers as a keynote speaker and honorable guest for several international conferences in cancer and diabetes as well as other recent biomedical applications as stem cells and nanotechnology.
Cancer is the number one killer in developed countries. Treatment is based on the classical triad: surgery, chemotherapy and radiotherapy but new care and research is needed using several new original approaches including gene-therapy and immune therapy. Based on our previous research and expertise, we developed a new approach of cancer treatment associating: 1) the sensitization of tumors to a pro-drug (anti-cancer drug not directly toxic for cancer cells but transformed into toxic metabolites) thanks to a suicide-gene, 2) the triggering of a secondary immune response directed against tumor cells. We built and patented an optimized suicide-gene able to transform cyclophosphamide (CPA) very efficiently into toxic metabolites (PCT/EP2012/058219, and US-2014-012718A1). Needing a vector, we used mesenchymal stem cells (MSCs), in mouse tumor models. Injection into the tumor of MSCs expressing our suicide gene followed by the injection of CPA resulted in a complete eradication of tumors in 30% of mice without recurrence more than 18 months after any treatment. Moreover, re-challenge experiments in cured mice demonstrated an immune response against tumor cells that helped to prevent recurrence and the onset of metastasis These encouraging results could nevertheless be improved by controlling the implantation of MSCs near the tumor. We demonstrated, in an orthotopic model of hepatocarcinoma (VX2) in rabbits, that intra-arterial injection of transduced murine MSCs entailed a decrease in tumor size, and, in some cases the non-occurrence of metastases. This method of injection is rapidly expanding in human clinical practice. It can be repeated several times and provides access to a large number of tumors. In humans, the isolation and multiplication of GMP quality CSMs is carried out in different laboratories and given the low immunogenicity of MSCs, transduced MSC clones, which are particularly effective for metabolizing CPA could be injected into different individuals (allogeneic). We also demonstrated, in vitro, the efficacy of transduced MSCs containing the suicide gene on a large number of human tumor cells (colon, breast, ORL…). Human hepatocarcinoma is a cancer with a survival rate of less than 20% at 5 years, and will be our first target but this strategy could be extended with a similar efficiency to all solid tumors.
She has completed her Ph.d in Molecular Pharmacology, Experimental Pharmacology and Metabolism, Physiology of Nutrition. She is the member of administration council of the Faculty of Medicine in Paris Descartes University, member of the American Association for Cancer Research. She has been appointed as the Mayor of Lignières en Vimeu from 1995, President of the Community of Communes of the Region of Oisemont from 2014,Departemental councilor from 2014. Her special area or research are Xenobiotic metabolism, Toxicology, Molecular Biology, Cancer, Resistance to chemotherapy, Gene therapy, Transcriptomic, Metabonomic.
Vitamin D3 potentiate the effect of 5-flourouracil on hepatocellular carcinoma induced in rats Laila A Eissa a,*, Amro EL-Karef b , Mohamed A Elmesery c, Amal R Ebrahim a a,c Department of Biochemistry, Faculty of Pharmacy, Mansoura university, Mansoura, 35516, Egypt b Department of pathology , faculty of medicine, Mansoura university, Mansoura, 35516, Egypt Abstract Hepatocellular carcinoma (HCC) ranks as the fifth commonest malignancy worldwide. Therefore, we aimed to study the role of vitamin D3 in treatment of hepatocellular carcinoma either alone or in combination with 5-flourouracil and the molecular mechanism of vitamin D3 action in the treatment of hepatocellular carcinoma. HCC was induced in SD rats by thiocetamide (200 mg/Kg). Rats were divided into 5 groups: control, HCC, HCC treated by 5-FU (5-FU), HCC treated by vitamin D3 (Vit. D3), HCC treated by 5-FU+vitamin D3 (5-FU/D). liver impairment was assessed by measuring serum α-fetoprotein and investigating liver sections stained with H/E. quantitative RT-PCR was used to measure the gene expression of caspase-3, transforming growth factor β1 (TGF-β1 ) and nuclear factor-erythroid 2 related factor 2 (Nrf2) was measured, while the immunohistochemistry was used to measure the expression of caspase-3 and TGF- β1. Vitamin D/5-FU co-therapy significantly decreased the expression of TGF-β1 and Nrf2 (P < 0.05 ) also significantly decreased serum AST, ALT, GGT, ALP activity, serum total bilirubin and AFP in comparison with their corresponding monotherapy, they also significantly increased serum albumin and expression of caspase-3. vitamin D3 combined with 5-FU exhibited a significantly greater anti-cancer effect than that of vitamin D3 or 5-FU treatment alone and thus combined therapy may present a novel therapeutic regimen for HCC. Further study is required to determine the molecular mechanism by which combined treatment with vitamin D and 5-FU inhibits the growth of HCC. Keywords: Hepatocellular carcinoma; vitamin D3; 5-flourouracil
Abiogenic evolution of aminoacyl-tRNA synthetase Kozo Nagano Nagano Research Institute of Molecular Biology, 4-8-24 Higiriyama, Kohnan-ku, Yokohama 233-0015, Japan The origin of life on earth demands a prerequisite for abiogenic evolution of RNA which is believed to be highly destructive on the earth covered by boiling ocean. I have proposed a mechanism of evolution of RNA as a left-handed single-stranded lattice with a trend of condensation and dehydration of organic compounds under a condition of high pressure and high temperature in the mantle of the earth. All organic compounds and energy-rich polyphosphates were synthesized from a large amount of methane hydrate, inorganic phosphate and oxygen radicals that were produced from ferrous oxides by disintegration of uranium. The six fold symmetrical structure of the lattice allowed to stack a lot of energy-rich compounds such as adenosine triphosphate and phospho-lipids which were required for formation of cell membranere at the later stage. More importantly, the cavity formed inside the lattice allowed polymerization of small L-amino acids such as glycine, alanine, proline and serine that were required to protect RNAs from hydrolysis. The 6.5 fold symmetrical lattice allowed a little more larger L-amino acids to incorporate the protective proteins and formation of α–helices, but insertion of larger amino acids such as arginine and methionine were rather difficult. However, in the next stage of evolution of the lattice formation of left-handed single stranded RNA, the 8 fold symmetric lattice seems to incorporate arginines and also the dimeric relationship between the two neighbor strands suggests a possibility of aminoacyl-tRNA synthetase to evolve because some of the aminoacyl–tRNA synthetases were found to be in dimeric form.
Kozo Nagano has completed his PhD from University of Tokyo, Pharmaceutical Sciences, Japan in 1963. He has published more than 40 papers in reputed journals. He has promoted to Professor of University of Tokyo at the Faculty of Pharmaceutical Sciences, in 1969.
Gliomas have intrinsic genetic traits allowing them to transform to more aggressive tumor types. This phenomenon makes the treatment of glioma extremely complicated and difficult. Among many transformation genetic aberrations is the overexpression and alternative splicing of the gene KCNMA1, which encodes the pore forming α-subunit of large-conductance calcium-activated voltage-sensitive potassium (BKCa) channels. KCNMA1 splicing is likely to play a regulatory role in glioma invasion, migration, and progression. In fact, we found a critical role for alternative pre-mRNA splicing events of KCNMA1 in invasion and proliferation of glioma cells, glioma growth, progression and diffusion. We have identified a hitherto unknown KCNMA1 mRNA splice variant with a deletion of 108 base pairs of exon 22 (KCNMA1vE22) mostly overexpressed in high grade gliomas, and elucidated the variant’s specific role in glioma biology. The KCNMA1 splicing effects and the potential role of KCNMA1vE22 as a critical posttranscriptional regulator of BKCa channel isoform expression is presented in this research paper. The resulting BKCa channel function in glioma is shown by expression and functional assays in cell culture and human glioma xenograft mouse models. The investigation of glioma tissues from brain tumor patients demonstrates that KCNMA1vE22 is a novel transformation biomarker of gliomas, and was mostly expressed in high-grade gliomas including glioblastoma multiforme (GBM, grade IV). Our study also implies that gliomas express KCNMA1vE22 and BKCa channel isoform to gain advantage in terms of aggressive growth and transformation. Targeting this variant could be a viable diagnostic, prognostic and treatment strategy as demonstrated by significantly attenuated growth of tumors in mice injected with KCNMA1vE22 cell lines expressing short-hairpin RNA (shRNA). Ultimately targeting the KCNMA1 variants will be clinically beneficial strategy to attenuate glioma transformation to an untreatable high grade glioma.
Synergistic effects of shikonin on the hyperthermia-induced apoptosis and its molecular mechanism De-Jun Zhou1, Jin-Lan Piao2, Yu-Jie Jin3, Meng-Ling Li3, Lu Sun3, Shahbaz Ahmad Zakki3, Qian-Wen Feng3, Hidekuni Inadera3, Zheng-Guo Cui1, 3* 1Henan Polytechnic University, Jiaozuo 454000 China; 2Department of Radiological Sciences, 3Department of Public Health, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan ABSTRACT Hyperthermia (HT) has been widely used for cancer therapy, however, it often fails to induce tumor cell killing sufficiently. Therefore, developing a non-toxic enhancer for HT-induced cell death is a critical hyperthermic cancer therapy. Shikonin (SHK) is a natural naphthaquinone derivative from a Chinese herbal medicine. In our previous study, we have reported that SHK is able to induce apoptosis and necroptosis in some cancer cells. In this study, the synergestic effect of SHK in low doses for mild HT induced apoptosis in the human myelomonocytic lymphoma cell line U937 was investigated. The cells were treated with HT at 44ºC for 10 minutes with or without SHK pre-treatment at low doses of 0, 0.1, 0.2 and 0.5µM. The results revealed that 0.5µM SHK significantly enhances HT-induced apoptosis as indicated by DNA fragmentation, caspase-3 activation, increased generation of intracellular reactive oxygen species (ROS) and lowering of mitochondrial membrane potential (MMP). It was also found that pro-apoptotic proteins like tBid, as well as Noxa, were remarkably expressed; while, expression of anti-apoptotic proteins including Bcl-2, Bcl-xL and Mcl-1 were decreased under the combined treatment. Also, observed phosphorylation of JNK and PKC- δ, and dephosphorylation of ERK and AKT in the combined treatment, that may have compounded the induction of apoptosis. In conclusion, SHK enhances HT-induced apoptosis via mitochondria-caspase dependent pathway. Its underlying mechanism involves increasing the intracellular ROS generation, inducing mitochondrial dysfunction, and activating PKC-δ and JNK.
I, Dr. Zheng-Guo Cui graduated from College of Medicine, YanBian University, China (1990-1996) and obtained my Ph.D. at Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan (2001-2004). Then I completed Postdoctoral Fellowship of Japan Science and Technology Agency focusing on various cell death mechanisms (2006-2007). Since 2009 I am working as an Assistant Professor in the Department of Public Health, Faculty of Medicine, University of Toyama, Japan. My research areas of expertise include redox signaling and oxidative stress, molecular mechanism of various cell death, sensitization of cancer therapy and influence of environmental pollutants on cancer risk. I have more than 30 publications in international reputed journals and received an International Award for Advanced Study by Young Scientist from International Association for the Sensitization of Cancer Treatment (IASCT) in 2005. Awarded the “Prize for Encouragement by The Japanese Society of Hygiene” from the Japanese Society for Hygiene in 2015.
Skin cancer is the most common type of cancer in Brazil. Among the different forms of presentation, cutaneous melanoma is distinguished by its severity. Melanomas are highly metastatic tumors, and its lethality is higher. Previous studies directed by our team at UFPR demonstrated that M1 may act modulating the activity of immune cells. In addition, through direct action or through modulation of the immune response, results also point to an antitumoral action in different mice and human cell lines showing significant changes. Thus, this study aims to evaluate the action of M1, natural complex highly diluted, on the human melanoma tumor cells as well as immune cells interaction. Human metastatic melanoma cell line 1205Lu and mononuclear cells isolated from human periphery blood mononuclear cells by leukophaeresis filters system after apheresis procedure were used. It was observed that the culture of melanoma cells in the M1 treated group had lower cell density due to cells death and to proliferation reduction. Human mononuclear cells after in vitro treatment and subsequent co-cultured with 1205Lu cells, observed at scanning electron microscopy, also showed a lower cell density, with images of tumor cells undergoing apoptosis. Markers were used to demonstrate the reduction of the number of viable cells (PI, annexin V, β-catenin). All they showed significant difference when compared with control group. After previous treatment of the mononuclear complex with M1 and subsequent co-cultivation for 48 hours with 1205Lu cells, there was also a significant great number of NK cells (p <0.0001), with a significant increase in number of activated NK cell. The results achieved in this work signal to M1 as a possible active tool with adjuvant potential of the therapy already available.
Dorly de F. Buchi is senior research at Molecular and Cell Biology Post Graduation at Federal University of Paraná, Brazil. Published many works about natural products modifyers of the biological response, immune system and mice and human tumor cells, in vitro and in vivo, using electron microscopy, cell culture, flow cytometry, and histopatological scanning microscopy
SEMINOMA: AN UNTAPPED RESERVOIR OF CANCER/TESTIS ANTIGEN SPECIFIC T CELLS FOR IMMUNOTHERAPY ABSTRACT Ika Nurlaila, Hayden Pearce, Suzy Eldershaw, Paul Moss Cancer/testis antigens (CTAgs) are of immense interest for use in immunotherapy. They are expressed only in cancers, not in normal tissues with the exception of the testis and, recently, they were also reported to be expressed in placenta. The antigens do not induce T cell-mediated immune response in testis and placenta since testis and placenta are immune privilege sites. This study focused on utilizing seminoma, a type of testicular germ cell tumor (TGCT) as an untapped reservoir of CTAgs-specific T cells. The evaluated CTAgs were MAGE-A1/A3/A4 and PLAC-1. A response was considered positive if the effector cytokine secretion (TNF-α and/or IFN-γ) exceed 0.1% after normalised. A potentially novel HLA-B44-restricted peptide from MAGE-A4 was previously found in seminoma. NetMHC, an online algorithm program, predicted the peptide to bind allele B44 weakly, however, it is encouraging to investigate. In addition to peptide-screening, global immunophenotyping was also performed to investigate the immune cell composition of the potential CTAg sources as well as immune checkpoint composition of the T cell subsets. Preliminary data suggests TIGIT and PD-1 may be important checkpoint markers in the cancer. Early results also indicate the proportion of MAIT cells may differ between nonseminoma and seminoma patients. This comprehensive understanding would be of importance to map CTL-mediated immune responses against CTAgs. Keywords: Checkpoint, CTAgs, immune response, MAGE-A, seminoma, PLAC-1
Background: Several diseases have been caused by contamination of surface and groundwater. Aim: The aim of the present work is to investigate the impact of iron overload in drinking water on liver pathology. Materials and Methods: Samples of drinking water, blood and true cut liver biopsies were taken from selected inhabitants. Those inhabitants (16 patients) from Mit-ghamr and Aga districts were suffering from liver disorders (had hepatitis C) and four patients had chronic cholecystitis from Mansoura district as control cases in relation to the type of drinking water. Samples of water, blood and true cut liver biopsies were taken and undergone for determination of iron level. Liver biopsies were taken routinely from patients group (16) and from control group (4) after having informed consent and during cholecystectomy. Measurement of iron level in water samples was carried out in duplicate with the use of GBC atomic absorption spectrophotometer, Taco company (Australia). Analyzed for serum iron level with a micro lab 200 spectrophotometer by using Iron-B kit, Biocon company (Germany). Results: The mean value of iron in surface water is lower than the permissible limit of Egyptian ministry of health (EMH) and World health organization (WHO). However, the mean value of iron in groundwater samples is higher than that permissible limit and than those of surface drinking water. Comparison between iron level in drinking water and human blood samples shows positive relationship. The control group depended on drinking surface water and had normal liver function tests, whereas the patient group that depended on drinking groundwater had abnormal values in liver function tests. These data suggest that the polluted iron drinking water is the reason for the liver disorder of the patients. Siderosis was apparent among those patients drinking polluted iron water in comparison to control cases. The siderosis appears to be responsible for resistance to treatment of HCV and progression of fibrosis. Conclusion; The accumulation of iron in liver leads to fibrosis. Iron depletion therapy could interfere with fibrosis development and possibly reduce the risk of hepatocellular carcinoma (HCC). Keywords; Water, blood, liver, Iron
Guidelines for Stem Cell Donor Selection,Which One is Better? Rabab Al Attas, MD, ABHI, ABMLI Historically, the best outcomes of allo-HSCT have been obtained when the donor is an HLA-matched sibling. Unfortunately, each sibling of a patient has only a 30 %chance of being HLA-matched, and with the small family sizes seen patients might have even less chance of having identical sibling. For those who do not have identical sibling; unrelated matched (MUD) allo Hematopoietic cell transplantation (HCT) from adult unrelated donors and unrelated umbilical cord blood (UCB) units can cure patients with malignant and nonmalignant hematologic diseases who lack identical family member donor. One prerequisite for hematopoietic stem cell (HSC) transplantation is a source of stem cells selected to match the patient’s HLA types to reduce the risks of allorecognition which leads to graft rejection, graft vs. host disease, and transplant-related mortality; and to increase the likelihood of immune reconstitution. Registries of unrelated volunteer donors and umbilical cord blood banks are sources of potentially matched stem cells. Selection of a suitable graft for allogeneic hematopoietic stem cell transplantation involves consideration of both donor and recipient characteristics. Of primary importance is sufficient donor recipient HLA matching to ensure engraftment and acceptable rates of GVHD. Nevertheless, well matched donors cannot be found for many patients, and many other patients either relapse or become too ill while waiting for a suitable donor to be identified. In this Perspective, the National Marrow Donor Program and the Center for International Blood and Marrow Transplant Research provide guidelines, based on large studies correlating graft characteristics with the clinical transplantation outcomes. The diversity of the HLA system complicates the search process and presented many challenges to the identification of best donors, this requiring sophisticated registry algorithms for matching, and expertise in allele and haplotype frequencies and associations to design search strategies. The development of accurate and high resolution DNA-based HLA testing methods and the continuing characterization of allele and haplotype frequencies have been powerful tools in donor search. Selection of the best available adult donor or UCB unit is necessary to maximize the likelihood of successful HCT. Therefore improved understanding of the importance of HLA matching at the allele level for HLA-A, HLA-B, HLA-C, and HLA-DRB1/ DQB1 loci, will lead to significantly improved outcomes of MUD transplantations. This review will discuss how should the best mismatched donor be selected? The presentation will also go through the effect of various HLA-typing methodologies and search strategies for the identification of the best donor from HLA- prospective. Recent results from a NMDP analysis will be also summarized.