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Nicb Research Overview

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The NICB (National Institute for Cellular Biotechnology) is located on the Dublin City University (DCU) campus in Dublin, Ireland. It is a leading multidisciplinary centre of translational research in fundamental and applied cellular biotechnology, molecular cell Biology, ocular diseases and biological chemistry. It includes a multidisciplinary team of Cell and Molecular Biologists, Biotechnologists, Chemists and Informatics specialists.
The NICB prioritises translational research involving collaborations with industry and with clinicians, and is committed to educating people from all backgrounds in the area of Biomedical Science.
This slideshare summarises the main research areas of the NICB, including:
Molecular basis for biopharmaceutical production by animal cells
Cancer – drug resistance, invasion and biomarkers
Tissue Engineering/Stem Cell Therapy – ocular diseases, diabetes
Using animal cells as research tools and models for disease research

Published in: Health & Medicine
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Nicb Research Overview

  1. 1. N a t i o n a l I n s t i t u t e F o r C e l l u l a r B i o t e c h n o l o g y Research Overview
  2. 2. NICB – Dublin City University • Government-designated National Centre of Expertise in Basic and Applied Molecular Cell Biotechnology since 1987 • New 3,200m2 building (opened 2006)
  3. 3. Main Areas of Research • Molecular basis for biopharmaceutical production by animal cells • Tissue Engineering/Stem Cell Therapy – ocular diseases, diabetes • Cells as research tools and models for disease research • Cancer – drug resistance, invasion and biomarkers
  4. 4. Cells as products Stem cell therapy/ Tissue engineering
  5. 5. Tissue Engineering/ Stem Cell Therapy • Twin focus – basic research and clinical application • Corneal stem cell transplant – entering clinical phase • Pancreatic islet transplant – currently technology transfer stage (Partner: University of Oxford)
  6. 6. • The Corneal surface consists of an epithelial layer that, in the healthy eye, is constantly renewed. • Located at the limbus a narrow ring of stem cells surrounding the cornea. Stem Cell Treatments For Eye Injuries 1
  7. 7. Stem Cell Treatments For Eye Injuries 2 • Patients with limbal deficiencies are unable to maintain a stable cornea. e.g. Chemical and thermal burns, Stevens-Johnson syndrome • Cornea transplantation in these patients typically fail due to conjunctival invasion, vascularization, and persistent epithelial defect. • The NICB in collaboration with its partners, proposes to introduce a stem cell therapy for these patients.
  8. 8. • Isolate and culture stem cells from the limbal ring. These cultured cells are then transplanted on to the surface of the injured eye, repopulating the damaged cornea • Research to refine techniques, find better differentiation markers and investigate biology of the process, e.g. using microarrays and proteomics Stem Cell Treatments For Eye Injuries - 3
  9. 9. Diabetes Research at the NICB
  10. 10. 5-10% of these have severe recurring hypoglycemia 23,000 adults in Ireland suffer from Type 1 Diabetes (T1DM)
  11. 11. Significant sustained decrease in severe hypoglycaemic events and improved quality of life. Islet transplantation
  12. 12. Islets from the Donor’s pancreas are infused into the recipient’s hepatic portal vein and produce insulin
  13. 13. Islet Transplantation Islet Isolation & Purification Donated Pancreas
  14. 14. DRIVE – Horizon 2020 funded project Diabetes Reversing Implants with enhanced Viability and long-term Efficacy
  15. 15. Mitochondrial Function Oroboros oxygraph Measures oxygen consumption in cells and tissues in real time
  16. 16. Cells as factories Manufacture of recombinant proteins
  17. 17. Research Related to Biopharmaceutical Production Our goal is to work closely with biopharmaceutical companies to improve basic understanding of the molecular basis for recombinant protein production by mammalian cells (including CHO and hybridomas) and • to translate this understanding into improved biopharmaceutical production processes • Special focus on miRNA
  18. 18. Why do we need improved biopharmaceutical production processes? • Increasing number of products • Current high unit cost • Reduced production costs will contribute to (a) Access for patients to treatments they need (b) Long-term health of the biopharmaceutical industry
  19. 19. Invited talks
  20. 20. Cells as research tools & as models for disease research
  21. 21. Cell culture Access to living human material - primary cultures - cell lines
  22. 22. Cell culture systems may not always completely reflect complex multi-cell type and spatial organisation in the body
  23. 23. C o r e t e c h n o l o g i e s : M o l e c u l a r p r of i l i n g of c e l l u l a r r e s p o n s e e . g . t o d r u g s ; t o t e m p e r a t u r e c h a n g e : t o o x i d a t i v e s t r e s s • mRNA (Affymetrix) • miRNA (ABI) • Proteins • Importance of bioinformatics analysis
  24. 24. Proteomics • Proteomics Infrastructure at the NICB • 2D DIGE (Difference Gel Electrophoresis) • Mass Spectrometry for Protein Identification/Characterisation, Including phosphoproteomics • Quantitative LC-MS approaches (i.e. gel-free)
  25. 25. Mass Spectrometry Suite • LTQ Orbitrap XL (Thermo Fisher Scientific) High mass accuracy Interfaced with Dionex Ultimate 3000 1D and 2D LC capabilities • LTQ XL with ETD Electron Transfer Dissociation (ETD) PTM analysis (e.g. phosphorylation) • MALDI TOF-TOF 4800 (Applied Biosystems) High throughput MALDI MS/MS LC-MALDI
  26. 26. Using proteomic profiling to identify new cancer markers in blood
  27. 27. Cancer Biomarkers for What? • Early detection • Monitoring of tumour burden (response to therapy) • Detection of relapse • Predicting response to specific therapy
  28. 28. Advanced Lung Cancer Haptoglobin ELISA 0 1000 2000 3000 4000 5000 6000 ug/mL Control Squamous Adenocarcinoma Small cell carcinoma
  29. 29. Ultimate aims of our Cancer Research Programmes Paired diagnostic and therapeutic approaches to treatment of individual cancer patients
  30. 30. Toxicity Testing Drug Resistance in Cancer Mechanisms of Cancer Cell Invasion
  31. 31. CHEMOTHERAPY WORKS, BUT ONLY SOMETIMES: WHY? One reason is drug resistance (acquired or intrinsic)
  32. 32. Adriamycin Distribution in Resistant Cancer Cells DLKP-A resistant cancer cells after exposure to Adriamycin. Fluorescent view DLKP sensitive cancer cells after exposure to Adriamycin Fluorescent view
  33. 33. Immunodetection of MRP-1 MRP-1 Negative Breast Carcinoma MRP-1 Positive Breast Carcinoma
  34. 34. Cancer invasion & Metastasis • Cell models, including clonal variants • In vitro properties and investigation of relevance in human cancer • Molecular profiling • New antigens/monoclonal antibodies
  35. 35. Therapeutic Targeting Using ADCs
  36. 36. Identification and Investigation of novel membrane protein targets with high cancer specific expression for potential therapeutic targeting using Antibody Drug Conjugates (ADCs)
  37. 37. Colon Cancer The 9E1 target antigen is highly expressed in colon cancer and show limited expression in normal colon
  38. 38. Pancreatic Cancer Novel membrane target is highly expressed in Pancreatic Cancer
  39. 39. Breast Cancer Novel Membrane Target is highly expressed in triple negative breast cancer (TNBC) and shows very limited expression in normal breast
  40. 40. Development of functional anti-invasive MAbs to identify potential drug targets associated with cancer invasion/metastasis.
  41. 41. MAb 7B7 targeting the Ku70/80 heterodimer significantly blocks the invasion of MiaPaCa-2 pancreatic cancer cells
  42. 42. Effect of LY6G6F siRNA on 2D colony formation of Mia Paca-2 cells Cell number seeded: 100 200 400 Control Negative siRNA LY6G6F siRNA #2
  43. 43. Uveal Melanoma
  44. 44. Proteomic Analysis of uveal melanoma to understand metastatic disease
  45. 45. Enucleation & Plaque Radiotherapy Treatments for Primary Tumor
  46. 46. metastasis in 50% of cases Primarily to the liver with average survival of 5-8 months
  47. 47. Loss of chromosome 3 Class I v Class 2 gene signature Loss of BAP1 GNAQ/GNA11 mutation Current Biomarkers
  48. 48. Aims of t h e St u dy : Iden tif y differen tially expressed protein s in primar y u veal melan oma t issu es of pat ien t s wh o developed met ast at ic disease ver su s t h ose wh o did n ot . Un der st an d t h e biology of t h e disease Iden tif y n ew th erapeu tic targets
  49. 49. Cancer Biotherapeutics
  50. 50. Background • NSABP-B31 (Paik et al. 2007) and N9831 (Perez et al. 2010) • Could a strong immune response to trastuzumab in the adjuvant setting be responsible for the response in HER2 low patients? • Trastuzumab has two mechanisms of action 1) it inhibits HER2 signaling and 2) engages the immune system through ADCC.
  51. 51. Antibody-Dependent Cell- mediated Cytotoxicity ADCC
  52. 52. NK cells and peripheral blood mononuclear cells (PBMCs) Immune cells from healthy Volunteers
  53. 53. Immunoblotting and high content analysis. HER2 Levels quantified
  54. 54. Examine the effects of TKIs on HER2 expression in a HER2- positive cell line (SKBR3) Laser scanning Confocal Microscopy
  55. 55. Determining the effect of PD-1/PD-L1 inhibition on response to trastuzumab in preclinical HER2- expressing breast cancer models.
  56. 56. Assessing the expression of immunomodulatory proteins in HER2-targeted therapy-resistant tumour cells.
  57. 57. Pancreatic Cancer Programme
  58. 58. • St Lukes Radiation Oncology Network, Rathgar • University of Buffalo Collaboration
  59. 59. Generation of primary pancreatic cancer and stromal cell lines
  60. 60. With pancreatic cancer cells and how they impact treatment success Stromal Cancer Cell interactions
  61. 61. L e v e l o f c o l o ny fo r m a t i o n i n Pa n c - 1 c e l l s p o s t c o - c u l t u r e w i t h p a n c re a t i c p a t i e n t - d e r i v e d f i b ro b l a st s .
  62. 62. Medicinal Inorganic Chemistry
  63. 63. artificial metallonuceases • First reported “self-active” oxidative system capable of inducing single-stranded DNA scission in the absence of exogenous reductant or oxidant. • Copper based complex, di-nuclear structure. • Displays excellent in vitro chemotherapeutic activity toward cisplatin-resistant ovarian cancer.
  64. 64. • Detection and quantification DNA oxidation by synthetic artificial metallonucleases using capillary electrophoresis. • Analysis completed using Agilent Bioanalyzer 2100 located within the molecular biological laboratory at NICB. Chemical nuclease detection using microfluidics
  65. 65. • Potent non-covalent DNA binding agents where nucleic acid recognition is achieved through use of the “phosphate clamp”. • Phosphate clamp-DNA interactions result in condensation of superhelical and B-DNA. • Triplatin-DNA binding inhibits endonuclease activity by type II restriction enzymes. • High chemotherapeutic potential for human cancer. Novel chemotherapeutic platinum(II) complexes
  66. 66. • Detection of apoptosis induced mitochondrial depolarization through exposure to ROS active copper developmental therapeutics. • Flow cytometry and confocal imaging analyses. • Broad spectrum of activity identified using the National Cancer Institute (NCI) 60-human cancer cell panel. • Unique mechanism compared to marketed therapeutics. Copper chemotherapeutic drug development
  67. 67. • Individual morphine molecules and derivatives thereof lack nucleic acid recognition properties. • In the triplet drug form unique properties emerge with this alkaloid substructure interacting with dsDNA and condensing superhelical DNA. Tripodal opioids as unique DNA interacting agents
  68. 68. Phenazine based biomaterials • Enhanced high affinity DNA binding • Distinctive nucleotide binding specificity. • High intercalative capacity • In vitro chemotherapeutic potential
  69. 69. • Metal catalyzed reactive oxygen species (ROS) in biological systems can cause a wide variety of pathological conditions including cancer. • The extent of DNA damage owing to these radicals can be quantified through 8-oxo-2’- deoxyguanosine (8-oxo-dG) lesion detection using both ELISA or LC-MS/MS analysis. Detection of free radical DNA damage
  70. 70. Metallodrug topoisomerase inhibitors • Development of intercalating metal complexes to unwind and relax negatively supercoiled DNA. • Applications as unique topoisomerase inhibitors.
  71. 71. Collaboration with Industry • Cells as factories • Cells as products • Cells as Research Tools • Cells for toxicity assay and bioassay • Making monoclonal antibodies • Whole genome mRNA and miRNA profiling • Proteomic profiling • Pharmacokinetics/drug analysis
  72. 72. Some past & present industry collaborators
  73. 73. Questions? N a t i o n a l I n s t i t u t e f o r C e l l u l a r B i o t e c h n o l o g y D u b l i n C i t y U n i v e r s i t y, G l a s n e v i n , D u b l i n 9 , I r e l a n d . + 3 5 3 1 7 0 0 5 7 0 0 w w w. n i c b . i e n i c b @ d c u . i e

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