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PolExGene

PolExGene. 18 Months Midterm Meeting Ghent, 17-18 / 12 / 2007. WELCOME IN GHENT. Sixth Framework Programme Priority 1 Life Sciences, Genomics and Biotechnology for Health. PolExGene

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PolExGene

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  1. PolExGene 18 Months Midterm Meeting Ghent, 17-18 / 12 / 2007 WELCOME IN GHENT

  2. Sixth Framework Programme Priority 1 Life Sciences, Genomics and Biotechnology for Health PolExGene Biocompatible non-viral gene delivery systems for the ex vivo treatment of ocular and cardiovascular diseases with high unmet medical need Contract number: LSHB-CT-019114 Coordinator: Prof. Dr. E. Schacht (UGent) Scientific/Technical Supervision: Prof. P. Dubruel 18 Months Midterm Meeting Ghent, 17-12 / 12 / 2007

  3. AGENDA 18 MONTHS MEETING Tuesday morning 08.30 Welcome (E. Schacht) 08.35 Brief overview of project objectives (E. Schacht) 08.45 Presentation of WP1 Selection of CIP and CPP 08.45: IBMT 08.55: ENS 09.05 Presentation of WP2 Development of CPP-containing polymers 09.05: UGent 09.15: CNRS 09.25 Presentation of WP3 Development of CIP-containing polymer membranes 09.25: UGent 09.35: CNRS 09.45 Coffee break

  4. Tuesday morning 10.05 Presentation of WP4 Preparation of plasmids and CPP-containing polyplexes 10.05: UGent 10.15: UH.FP 10.25: IBMT 10.35: UKU 10.45 Presentation of WP5 Characteriation of polyplex-cell and polymer membrane-cell interactions 10.45: UH.FP 10.55: IBMT 11.05: ENS 11.15: UKU 11.25 Presentation of WP6 Study on immunological properties of polyplexes and polymer membranes 11.25: IMIC 11.35: UAT 11.45 Presentation of WP7 Polymer membrane implantation in test animals 11.45: UAT 12.00 Lunch

  5. Tuesday afternoon 13.30 Presentation of WP8 : management structure, progress and planning 13.40 Summary of the situation and review of the project technical objectives and time scale (E. Schacht and P. Dubruel) 14.40 Comments on the project status and project meeting Recommendations for the Mid-Term Report by the Project Officer : Dr. J. Sautter 15.20 Closing remarks and fixing dates and venues for next meetings (E. Schacht) Month 24 : Prague, 22-23 May 2008 Month 30: Montpellier, 13-14 November 2008 15.30 End of the meeting

  6. PARTNERS

  7. PARTICIPANTS UGent:E. Schacht, P. Dubruel, V. Vermeersch, S. Van Vlierberghe UH.FP:A. Urtti, A. Subrizi IBMT:H. Thielecke, H. Bueth IMIC:B. Rihova, M. Sirova ENS:A. Joliot, L. Tibaldi UAT:T. Peters, S. Julien ARK:Jani Raty (not present due to serious health problems) UKU:T. Wirth CNRS:J. Martinez, G. Subra, M. Amblard EU:J. Sautter (EC-coordinator)

  8. Strategy A: transfection cell seeding IMPLANTATION + cell CPP - based transfected polyplexes cell transfection Strategy B: + biodegradable cell - CPP - based interactive polymer polyplexes membrane PROJECT OBJECTIVES  Development of a non-viral gene therapy that will be applied for both ocular and cardiovascular diseases

  9. WORK PACKAGES Work packages: 1) Selection of CIP and CPP [month 1-28] 2) Development of CPP-containing polymers [month 1-32] 3) Development of CIP-containing polymer membranes [month 6-24] 4) Preparation of plasmids and CPP-containing polyplexes [month 1-36] 5) Characteriation of polyplex-cell and polymer membrane-cell interactions [month 1-36] 6) Study on immunological properties of polyplexes and polymer membranes [month 6-36] 7) Polymer membrane implantation in test animals [month 12-36] 8) Project management [month 1-36]

  10. WORK PACKAGES Work package 1:Selection of CIP and CPP (UGent, IBMT, ENS) The objective of WP 1 is to develop and select cell interactive peptides (CIP) and cell penetrating peptides (CPP) using a variety of techniques including impedance measurements, phage display and (molecular) cell biology techniques. The CIP will be selected for enhancing the cell interactive properties of electrospun polymer membranes, whereas the CPP will be selected for enhancing the internalisation and appropriate intra-cellular addressing efficiencies of polyplex formulations. Work package 2:Development of CPP-containing polymers (UGent, ENS, CNRS) The objective of WP 2 is to develop and characterise cell penetrating peptide (CPP), cationic polymers and their CPP-derivatives using different immobilisation strategies. Polymer conjugates with varying substitution degrees of CPP will be prepared to gain control over the cell penetrating properties of the macromolecules and the resulting polyplexes. A range of immobilisation strategies for immobilising peptides on polymeric gene delivery systems will be evaluated.

  11. WORK PACKAGES Work package 3:Development of CIP-containing polymer membranes (UGent, CNRS) The objective of WP 3 is to develop solvent-casted and electrospun non-woven nanofiber polymer membranes for cell seeding purposes. Different electrospinning parameters will be studied and optimised. In order to improve the membrane-cell interaction and to enhance the cell proliferation and differentiation, different cell interacting peptides (CIP) will be incorporated in the polymer membrane. Work package 4:Preparation of plasmids and CPP-containing polyplexes (UGent, UH.FP, IBMT, ENS, UKU) The objective of WP 4 is to develop therapeutic plasmids for the ocular and the cardiovascular aspect of the project. Plasmids with marker genes will be used for performing a detailed physico-chemical characterisation of cell penetrating peptide (CPP) containing polyplexes using a variety of techniques. Finally, the coating of polymer membranes and vascular grafts with CPP-containing polyplexes as well as the coating of vascular grafts with a polymer membrane will be studied in detail.

  12. WORK PACKAGES Work package 5:Characteriation of polyplex-cell and polymer membrane-cell interactions (UH.FP, IBMT, ENS, UKU) The objective of WP 5 is to study the interaction of different cell types, including retinal pigment epithelial (RPE) cells for the ocular application and vascular endothelial cells and smooth muscle cells for the cardiovascular application, with the polymer materials developed in WP 2 - WP 4. Both the interaction between CPP-containing polyplexes and cells and the interaction between CIP-containing polymer membranes and cells will be investigated. Work package 6:Study on immunological properties of polymers and polymer membranes (IMIC, UAT) The objective of WP 6 is to study possible immunological properties of the developed cell penetrating peptide (CPP) containing polyplexes and the cell interactive peptide (CIP) containing electrospun polymer membranes. Both the in vitro and in vivo immunological properties will be studied using state of the art techniques.

  13. WORK PACKAGES Work package 7:Polymer membrane implanatation in test animals (UAT, ARK, UKU) The objective of WP 7 is to implant the polymer membrane seeded with ex vivo treated cells in test animals. The technology developed will be applied for both ocular diseases (age-related macular degeneration) and cardiovascular diseases (stenosis and thrombosis in vascular grafts). Work package 8:Project management (UGent) The objective of WP 8 is to ensure proper administrative management, monitoring and control of the project and to ensure maximum progress towards the project objectives and its deliverables. In addition, the project management will also take care of innovation related activities (dissemination, exploitation and intellectual property rights).

  14. WORK PACKAGES Gantt diagram

  15. PRESENTATION • OF • WORK PERFORMED IN FIRST TERM • on a • WORK PACKAGE BASIS • Results achieved in first term • Planning for second term

  16. PROJECT MANAGEMENT Management board: Members:Prof. E. Schacht (P1), Prof. A. Urtti (P2), Dr. H. Thielecke (P3), Prof. B. Rihova (P4), Dr. A. Joliot (P5), Dr. T. Wheeler-Schilling (P6), Dr. D. Ellam (P7), Dr. T. Wirth (P8), Prof. J. Martinez (P9) Scientific/Technical Supervision Group Chairman: Prof. Peter Dubruel

  17. PROJECT MANAGEMENT Scientific/Technical Supervision group: Tasks: research follow-up  exchange of products among different partners Members:Dr. P. Dubruel (P1), Dr. M. Hornof (P2), Dr. H. von Briesen (P3), Dr. M. Sirova (P4), Dr. E. Dupont (P5), Prof. E. Zrenner (P6),Dr. D. Ellam (P7), Dr. T. Wirth (P8), Prof. J. Martinez (P9) Knowledge and Innovation Board: Tasks:dissemination of results intellectual property aspects exploitation of the project results Members: Prof. E. Schacht, Prof. A. Urtti, Dr. H. Thielecke, Dr. A. Joliot Advisory Board: Tasks: widespread dissemination of the scientific results/ activities to non-scientific organisations  providing advice to the PolExGene consortium Members:ESGT (European Society of Gene Therapy), EPPOSI (European Platform for Patients’ Organisations, Science and Industry, VIB (Flemisch Institute for Biotechnology), Prof. J. Kirkpatrick, Prof. Em. Berndt Ehinger, Prof. G. Fuhr, Qiagen

  18. MEETING AND REPORTING SCHEDULE Planned meetings: Management Board, Innovation Board and Scientific/Technical Supervision group meet every 6 months Reporting schedule: Internal report: every 6 months EU report: after 18 and 36 months

  19. MEETINGS • Technical meeting • Kick-off meeting Ghent, 23-23 Aug 2006 • Month 6 meeting Paris, 8-9 Feb 2007 • Month 12 meeting Helsinki, 23-24 Aug 2007 • Month 18 meeting Ghent, 17-18 Dec 2007 • Ad Hoc meetings • Paris, 17 Oct 2006 CIP & CPP peptides & cell transfection • Tubingen, 10 Dec 2006 polymer membranes • - Paris, 07 Sept 2007 in vitro transfection protocols

  20. CONSORTIUM AGREEMENT Finalised and signed by all partners end 2006 WEBSITE http://www.polexgene.eu

  21. Proposal for adjusted description of deliverables

  22. MILESTONES WP 1: M1-1: Availability of a (first series) of selected number of CPP that to be used for conjugation to DNA binding polymers and for incorporation in polyplex formulations (month 18). M1-2: Availability of a (first series) of selected number of CIP that will be used for enhancing the cell interactive properties of polymer membranes used for cell seeding purposes (month 18). WP 2: M2-1: Availability of a first selected number of cationic polymers and their CPP-derivatives that will be evaluated for their DNA binding properties and their biological properties (month 12).

  23. MILESTONES WP 3: M3-1: Availability of a first selected number of polymer membranes, based on biopolymers & CIP-containing biopolymers, that will be used for polyplex functionalisation and for biological studies (month 15). WP 4: M4-1: Availability of the first selected therapeutic and cell specific plasmids to be used for genetic guiding of cells seeded on polymer membranes used in transplantations (month 9). M4-2: First characterised polyplexes and CPP-containing polyplexes to be used in transfection and toxicity studies and for polymer membrane coating (month 12). M4-3: Availability of polymer membranes coated with polyplex formulations to be used for cell seeding purposes (month 20: +4 ).

  24. MILESTONES WP 5: M5-1: Availability of optimised CPP-containing polyplexes for transfection of retinal or vascular cells (month 24: +12). M5-2: Availability of CIP-containing polymer membranes on which polyplex-transfected retinal or vascular cells can be seeded, further grow and differentiate (month 28 : +8). M5-3: Availability of polyplex containing polymer membranes for cell guidance of retinal or vascular cells to be used for implantation purposes (month 30). WP 6: M6-1: The availability of a selected number of CPP-containing polyplexes and CIP- containing polymer membranes, with optimal immunological properties, that can be used for implantation in the retina or be used as coating for vascular prosthesis (month 32: + 8).

  25. MILESTONES WP 7: M7-1: The availability of polymer membranes seeded with therapeutic gene expressing cells that can be applied as implantation system for the treatment of age related macular degeneration (month 36). M7-2: The availability of polymer membranes on which therapeutic gene expressing cells are seeded, to be applied as coating for vascular prosthesis (month 36). WP 8: M8-1: Availability of the Consortium Agreement proposal that will be delivered at the latest during the kick-off meeting of the PolExGene project (month 1). M8-2: Availability of a report summarising the results obtained in the last 18 months (month 18 and month 36).

  26. WORK PACKAGES Gantt diagram

  27. Person months planned PolExGene midterm meeting Ghent, 17-18 / 12 / 2007

  28. BUDGET SPENDING

  29. Main reasons for underspending in first term • Recrutement of personnel dependent on first instalment • Difficulties (in Europe) to recrute skilled researchers • (particularly on the postdoc-level) : has caused significant delays • - Awareness that the project requires an introduction stage • for defining optimal requirements for synthetic gene • vectors, cell systems, in vitro cell transfection protocols • Awareness among the partners that the major work load would come in the • second term

  30. Comments and recommendations by Dr. J. Sautter

  31. Closing remarks and fixing dates and venues for next meetings Month 24 : Prague, 22-23 May 2008 Month 30: Montpellier, 13-14 November 2008 ‘Ad hoc’ meetings: planned 1) polymer membranes 2) polyarginine chemistry 3) IBMT, …

  32. THANKS Have a safe trip home Merry Christmas and Succesful 2008 Month 24 : Prague, 22-23 May 2008 Month 30 : Montpellier, 13-14 November 2008

  33. DELIVERABLES WP 1: D10: First Cell penetrating peptides (CPP) for polyplex formulations (month 18) D11: First Cell interactive peptides (CIP) for polymer membranes (month 18) WP 2: D2:FirstCationic polymer and cationic CPP-containing polymers as DNA binding vectors (month 12) STANDARD PROTOCOL FOR IN VITRO TRANSFECTION STUDIES WP 3: D5: Polymer membranes based on selected biopolymers and CIP-containing biopolymers (month 15)

  34. DELIVERABLES WP 4: D1: Cell specific and therapeutic plasmids (months 9) D3: Polyplexes and CPP-containing polyplexes (month 12) D6: Polyplex containing polymer membranes (month 16) WP 5: D4: Transfected retinal/vascular cells using polyplexes (month 12) D8: Transfected retinal/vascular cells seeded on polymer membranes (month 18) D12: Polyplex containing polymer membranes seeded with retinal/vascular cells using marker genes (month 24) D14: Polyplex containing polymer membranes seeded with retinal/vascular cells using therapeutic genes (month 30)

  35. DELIVERABLES WP 6: D9: In vitro and in vivo immunologically evaluated CPP-containing polyplexes (month 18) D13: In vitro and in vivo immunologically evaluated CIP-containing polymer membranes (month 24) WP 7: D15: Retinal cell seeded polymer membranes as implants for ocular diseases (month 36) D16: Vascular cell seeded polymer membranes as coating for vascular prosthesis (month 36) WP 8: D7: 18 month report D17: 36 month report

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