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About OMICS Group

About OMICS Group.

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About OMICS Group

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  1. About OMICS Group OMICS Group International is an amalgamation of Open Access publications and worldwide international science conferences and events. Established in the year 2007 with the sole aim of making the information on Sciences and technology ‘Open Access’, OMICS Group publishes 400 online open access scholarly journals in all aspects of Science, Engineering, Management and Technology journals. OMICS Group has been instrumental in taking the knowledge on Science & technology to the doorsteps of ordinary men and women. Research Scholars, Students, Libraries, Educational Institutions, Research centers and the industry are main stakeholders that benefitted greatly from this knowledge dissemination. OMICS Group also organizes 300 International conferences annually across the globe, where knowledge transfer takes place through debates, round table discussions, poster presentations, workshops, symposia and exhibitions.

  2. About OMICS Group Conferences OMICS Group International is a pioneer and leading science event organizer, which publishes around 400 open access journals and conducts over 300 Medical, Clinical, Engineering, Life Sciences, Pharma scientific conferences all over the globe annually with the support of more than 1000 scientific associations and 30,000 editorial board members and 3.5 million followers to its credit. OMICS Group has organized 500 conferences, workshops and national symposiums across the major cities including San Francisco, Las Vegas, San Antonio, Omaha, Orlando, Raleigh, Santa Clara, Chicago, Philadelphia, Baltimore, United Kingdom, Valencia, Dubai, Beijing, Hyderabad, Bengaluru and Mumbai.

  3. A proteomic-bioinformatic integrated approach for studying the effect of a peptide drug candidate on ovarian Cancer Maria Paola Costi Department of Life Science Unimore Modena

  4. Ovarian Cancer • Ovarian Cancer (OC) Itis the mostlethalof the gynecologicmalignancies • Non specific symptoms that accompany the early disease  OC diagnosis in advanced stage  therapeutic options are limited. • FIRST LINE TREATMENT • surgical cytoreduction + chemotherapy (and/or radiotherapy) • Chemiotherapy Platinum (cisplatin) /taxane (paclitaxel) combination • High initial response (≈ 70%) • CAUSES OF FAILURE • OC diagnosis in advanced stage (FIGO Stages IIB to IV) • Rapid occurrence of resistance to the standard systemic therapies • High percentage of relapse after 6 months (> 90%)

  5. Our aims are1. to develop new drugs against resistant ovarian cancer2. to provide a new methodology for fingerprint evaluation of biomolecules changes in cells3. to translate the concept to clinical samples

  6. Development of new drugs against resistant ovarian cancer

  7. Thymidylate synthase and folate pathway targeting

  8. Targeting Thymidylate synthase pathways hTS monomers hTS dimer active/inactive + ? mRNA DNA

  9. New strategiesagainstresistantovariancancer (ROC) • Pemetrexed (AlimtaTM) • Bindingat TS active site • Under clinicalevaluation for ROC • PMX(AlimtaTM)is an antifolatethatinhibitsTS, DHFR, GART and ATIC. • 14 clinical trials Pemetrexedphase II • Proteomicevaluation of the effect of PMX on: • OC celllines • OC biopsies from a Phase II clinical trial • Antifolate peptides • Binding at TS dimeric interface • Proteomic evaluation of the effect of peptides on: • OC cell lines • Primary tumor tissues • Cardinale D. et al.,PNAS, 2011, Pelà et al, J Med Chem.2014

  10. Can we approach the whole proteomic/targeted proteomic approach for a wider view of lead/drugs effect? Targeting the same target, different mechanism of inhibition Different cellular protein profile? Need for a more integrated non reductionistic approach

  11. Study of the effect of antifolate peptides in the cell lines through Proteomic-bioinformatic approach A2780 A2780/CP IGROV1 Treated with LR, LR-DGln4 and PMX To define a Protein Profile: proteins expression could be informative of the effect of peptides on OC cell lines.

  12. Flow chart

  13. Differential proteomic and bioinformatic approach • NETWORK ANALYSIS OF DEPS: 64 out of 160 DEPs resulted connected • using interrogation through: • BioGrid, IntAct and Reactome Up – regulated Down – regulated Up/Down – regulated Functional neighbors Up – regulated Down – regulated • FOUR MAIN CLUSTER ARE MODULATED BY LR: • Cluster a ribosomal proteins. • Cluster b proteins of pyrimidineribonucleotide metabolic process, • proteins RNA-connected and protein transport-connected. • Cluster cribonucleoproteins. • Cluster d proteasome complex. Up/Down – regulated

  14. Folate Pathway Members connected to DEPs Up–regulated Down–regulated Up/Down–regulated Folate–related PMX-related Physical protein interactions Lecture protein interactions 35 proteins from the original dataset and 14 folate-associated proteins

  15. ProteinsPanelSelection 4 proteins known to be directly involved in the folate cycle 3 folate related protein neighbor to proteins deregulated by LR treatment 3 differentially expressed proteins DEPs

  16. Western Blot targeted approach and panel validation

  17. How can we translate the profile in a easy read-out?

  18. Peptide L-Barcode A2780 peptide barcode LR 0 1 2 3 4 5 6 7 8 9 PROTEIN CODE: 0=GART, 1=TRAP1, 2=DHFR, 3=HSP90AA1, 4=ATIC, 5=MGMT, 6=TS, 7=MTHFR, 8=EIF2S1, 9=SHMT1

  19. LR treatment vs A2780 & IGROV1 treated with LR 5 μM (n=3) A2780 IGROV1 LR 0 1 2 34 5 6 7 89 LR 0 1 2 34 5 6 7 8 9

  20. LR effect in sensitive OC celllines Conserved code: 0 = GART 1 = TRAP1 2 = DHFR 3 = MGMT 4 = ATIC 5 = HSP90AA1 6 = TS IGROV1 peptide barcode IGROV1/A2780 peptide barcode A2780 peptide barcode LR 0 1 2 3 4 5 6 7 8 9 LR 0 1 2 3 4 5 6 7 8 9 LR 0 1 2 3 4 5 6 X X X CONSERVED code VARIABLE code

  21. LR-DGln4 treatment vs A2780 & IGROV1 A2780 peptide barcode IGROV1 peptide barcode LR-DGln40 1 2 34 5 6 7 8 9 LR-DGln4 0 1 2 34 5 6 7 8 9

  22. LR-DGln4 peptide L-barcode IGROV1/A2780 peptide barcode A2780 peptide barcode IGROV1 peptide barcode DGln4 0 1 2 345 6 7 8 9 DGln4 0 1 2 345 6 7 8 9 SIMILAR CONSERVED CODES SUGGESTING the SAME MECHANISM OF ACTION LR 0 1 2 3 4 5 6 X X X LR/LRDGln40 1 2 3 4 X X X X X

  23. Cisplatin resistant/sensitive OC cell lines A2780/CP peptide barcode LR-DGln4 0 1 2 3 4 5 6 7 8 9 A2780 peptide barcode SIMILAR CONSERVED CODES SAME MECHANISM OF ACTION A2780/CP LR-DGln4 0 1 2 X 4 5 X X X X A2780 LR-DGln4 0 1 2 3 4 X X X X X LR-DGln4 0 1 2 3 4 5 6 7 8 9

  24. Peptides L-barcode LR/LR-DGln4 0 1 2 3 X 5 X X X X X 0 = GART 1 = TRAP1 2 = DHFR 5 = HSP90AA1 X variable

  25. PTX barcode Vs LR-DGln4 barcode A2780 PTX barcode A2780 PTX/peptide barcode A2780 peptide barcode PTX 0 1 2 3 4 5 6 7 8 9 LR-DGln4 0 1 2 3 4 5 6 7 8 9 DIFFERENTS BARCODES DIFFERENT MECHANISM OF ACTION

  26. Heat map Statistical analysis of the WB results PCA. Different clusters for a different mechanism Heatmap. Blue down expression, red high expressing F.Genovese, MPCosti et al. J.Proteom Research, submitted

  27. What is the biological meaning of the protemic signature identified?

  28. Looking for a proteinsignatureinduced by LR…. OC A2780 CELLS CONTROL vs LR-peptide TREATED ..a proteomics-bioinformatics combined approach led to the identification of a set of proteins regulated upon treatment with LR-derived peptides. TRANSLATION OF THE PROTEOMIC RESULTS INTO BIOLOGICAL/MOLECULAR EFFECTS ON OVARIAN CANCER CELLS!

  29. DI-INACTIVE CONFORMATION ACTIVE CONFORMATION CK2 and other kinases P [DGln4] LR Ubiquitin-proteasome system Ribosomal proteins, RNA metabolism and translational initiation Stress-responsive pathways TRANSLATIONAL BLOCK eIF2A phosphorylation P BINDING TO SEVERAL CELLULAR mRNAs (TRANSCRIPTION FACTORS? RNA binding proteins/TRANSLATIONAL REPRESSORS? Stress-responsive proteins?) TRAP1 VDAC1 HSP90 -Opening of mitochondrial transition pore (MTP) -ROS increase APOPTOTIC CELL DEATH

  30. Conclusions • The results confirm that our new methodology provides: a) An approach to the discovery of molecular drug targets/off-targets which could be part of a pharmacodynamic profile to monitor candidate drugs activity since the very early phases of the drug development process. b) Deregulations can be mainly assigned to the following processes or functions: • regulation of translational initiation, • termination of RNA Pol-II transcription, • transport, • proteasome. 2. different expression level changes observed consequently to our peptides treatment in comparison with a folate antagonist currently used in therapy, Pemetrexed, are in agreement with the hypothesized different mechanism of action.

  31. Filippo Genovese Alessandra Gualandi Chiara Marraccini Silvia Pirondi Leda Severi Andrea Martello Domenico D’Arca Maria Rosaria Amoroso Gaetano Marverti Paul Perco Michela Pelà Remo Guerrini Glauco Ponterini Maria Paola Costi

  32. Thankyou.

  33. Let Us Meet Again We welcome you all to our future conferences of OMICS Group International Please Visit:www.omicsgroup.com www.conferenceseries.com www.pharmaceuticalconferences.com

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