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Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
Building Bridges Between Discovery, Preclinical, And Clinical Research 2008
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Building Bridges Between Discovery, Preclinical, And Clinical Research 2008

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Key translational research concept illustrated by personal experiences

Key translational research concept illustrated by personal experiences

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  • Dark teal text shows timing of sampling for B and T cell counts and for immune response
  • *say something in the title about the result, more thought on title Legend: T and B cell counts of individual patients enrolled in the 3M rest and 6M rest post rituximab cohorts. Horizontal bar represents the population median and the “whiskers” represent the inter-quartile range. Pre-R: cell numbers prior to first rituximab treatment (on average 9 days prior) Imm1: cell numbers immediately before the first immunization Imm4: cell numbers immediately before the fourth immunization Imm8: cell numbers immediately before the eighth immunization 8 Wks post: cell numbers 8 weeks after the eighth immunization Results: The absolute B cell numbers before rituximab treatment in both the 3M rest and 6M rest cohorts were generally low or even below normal for a significant numbers of patients, possibly due to the effect of prior chemotherapy treatment. Although patients enrolled in 3M rest and 6M rest cohorts were highly similar in term of age, gender, and disease status, the absolute B cells numbers before rituximab in the 3M rest cohort were generally lower than those in the 6M rest period. In both cases, B cell counts are expected to be essentially zero post rituximab. As expected, the absolute numbers of B cells after rituximab treatment were significantly suppressed (Imm1) and progressively increased with time (Imm4, Imm8, and 8 Wk Post) in both cohorts. The absolute T cells numbers before rituximab treatment in both the 3M rest and 6M rest cohorts were within normal ranges for the majority of patients. These numbers did not vary significantly throughout the immunization and the 8-week follow up.
  • There is a statistically significant difference between trial 2002-09, 3M and 6M Rest, and 9902B. There is also a significant difference between 2002-09 3M and 6M Rest periods.
  • Each B cell normally produces a unique B cell antigenic receptor (BCR, aka surface immunoglobulin, sIg) in order to give the cell immune specificity. The BCR defines the specific B cell as unique. NHL is a clonal population of tumor cells. The idiotype is the collection of unique antigenic determinants present in the variable region of the clonal immunoglobulin expressed by the B cell.
  • Transcript

    • 1. Cancer and Inflammatory Diseases Immunotherapy Building Bridges Between Discovery, Preclinical, and Clinical Research
    • 2. Outline
      • B Cell Lymphoma Immunotherapy
        • Active immunotherpay following rituximab treatment: from nonclinical study to the clinic
        • Improving on rituximab immunotherapy: from discovery research to preclinical model
      • Inflammatory Bowel Disease
        • Analysis of gene expression profile in patient’s biopsies: from clinical samples to basic research
        • Identification of a clinical biomarker from mode of action studies: from nonclinical research to clinical monitoring
      • Asthma
        • Analysis of anti-CD25 mAb mode of action in vitro: from nonclinical research to expanded clinical indication
    • 3. B Cell Lymphoma Immunotherapy
    • 4. B Cell Lymphoma
      • Follicular Non-Hodgkin’s Lymphoma
        • Clinical
          • Represents 25% of all cases of lymphoma in the US
          • Indolent malignancy: slow progression
          • Most patients will eventually relapse and will require more aggressive treatment
          • The duration of remission tends to decrease after each relapse
        • Pathology
          • Mainly localized to lymph nodes and the bone marrow
          • Malignant B cells preserve most of the features of mature B cells
            • Expression of surface Ig, CD19, CD20, CD21, and CD22
            • Propensity to organize into follicles
    • 5. Personalized Active Immunotherapy: concept
      • Indication
        • Follicular lymphoma
      • Drug
        • Unique vaccine for each patient
        • Based on unique Ig (idiotype) expressed by tumor
        • Linked to KLH  immunogenicity
      • Clinical regimen
        • Chemotherapy
        • Recovery (6 months)
        • Immunization
    • 6. B CELL LYMPHOMA IMMUNOTHERAPY
        • Active immunotherapy following rituximab treatment: from nonclinical study to the clinic
    • 7. Clinical Study Design Immunization efficiency following rituximab Rituximab 4 Doses (N=90)* Restage at 8 Weeks: CR, CRu, PR Patients who failed chemotherapy Study 2002-09 Sampling times for T and B cells and immune response *Third cohort (N=16) of immunized patients were not eligible for either 3M or 6M Rest groups. Screen failures (N=33) not eligible for immunization. Leonard J.P. et al., ASCO 2006 Follow- up 6-month Rest Period (N=23) 8 Immunizations Q 2 weeks Follow- up 3-month Rest Period (N=18) 8 Immunizations Q 2 weeks
    • 8. Patients treated with rituximab have dramatically reduced B cell numbers B Cells (CD19 + ) T Cells (CD3 + ) 3M rest period 3M rest period 6M rest period 6M rest period Leonard J.P. et al., ASCO 2006 Post-R Post-R
    • 9. Patients treated with rituximab have significantly lower humoral response against KLH Leonard J.P. et al., ASCO 2006 Analysis of variance (treatment effect) 9902B 2002-09 3M Rest p < 0.0001 9902B 2002-09 6M Rest p < 0.0001 2002-09 3M Rest 2002-09 6M Rest p < 0.05
    • 10. Conclusions
      • Rituximab treatment dramatically reduces the number of patient B cells
        • B cell numbers remained low up to 10 months after treatment
      • Humoral response in patients treated with rituximab is significantly reduced
        • 6-month recovery is not sufficient to restore a normal response
        • Significant improvement in response between 3- and 6-month recovery period
      • Adapt active immunotherapy clinical regimen
        • Allow for longer recovery time (if possible)
        • Schedule start of immunization in function of B cell recovery
      Include nonclinical exploratory end points to clinical studies  future clinical strategy
    • 11. B CELL LYMPHOMA IMMUNOTHERAPY
        • Can we improve on rituximab?
    • 12. B Cell Surface Targets CD22 CD20 CD19 BCR Idiotype  frequency ~1/10 10 V region framework  frequency ~ 1/20 C region  frequency ~ 1/2 CD19, CD20, CD22  frequency 1/1 Sornasse T. et al., ASH 2007
      • In Human
      • 45 Ig heavy chain V genes
      • 70 Ig light chain V genes
      • Ig V gene sub-families share common structures
      • Ig include 1 light and 1 heavy chain
    • 13. Concept of a selective mAb immunotherapy of B cell lymphomas Sornasse T. et al., ASH 2007
    • 14. mAbs specific for Ig variable region framework directly kill lymphoma cell line in vitro Anti-Ig VL mAb: Cell line A Anti-Ig VH mAb: Cell line B Lymphoma cell lines were incubated for 48 hours in the presence of mAbs. Dead cells were identified by flow cytometry as 7-AAD-positive cells.
    • 15. Main Hurdle in vivo: Soluble Antigen
      • Is it possible to deplete target B cells in vivo, in the presence of high level of soluble antigen ?
      Sornasse T. et al., ASH 2007 Anti-Ig mAb Serum Ig Target B Cell
    • 16. Incorporate Hypothesis Testing to Preclinical Study
      • Study type: Exploratory toxicology
      • Test article: anti-human Ig VH 3.23
      • Model: cynomolgus monkeys
        • Similar to human regarding frequency of target cells, levels of soluble antigen, and affinity for Ig sub-family
      • Study design
        • 3 Dose levels: 10, 40, and 100 mg/kg
        • 4 animals / group: 2 Males / 2 Females
        • Treatment regimen: 4 x q3-4 d
      • Exploratory end points
        • Frequency of target B cells and of total T and B cells
      Sornasse T. et al., ASH 2007
    • 17. Anti human Ig VH 3.23 mAb does not significantly affect the frequencies of B and T lymphocytes in vivo Control Group Dose Group: 10 mg/kg Dose Group: 40 mg/kg Dose Group: 100 mg/kg Legend Infusions Flow cytometry Schedule of events Sornasse T. et al., ASH 2007 Study Days 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
    • 18. Anti human Ig VH 3.23 mAb depletes target B cells in vivo (Study 2)          Sornasse T. et al., ASH 2007
    • 19. Conclusion
      • Monoclonal antibody specific for human Ig HV 3.23 can reach of target B cells despite the presence of soluble serum immunoglobulins
        • Method used to track target B cells can not differentiate between cell depletion and antigen down-modulation
      • In vivo administration of a mAb specific for human Ig HV 3.23 does not significantly affect the frequencies of T and B lymphocytes
      Challenge/verify original hypothesis throughout development Remain aware of model limitation and built-in assumptions
    • 20. Inflammatory Diseases
    • 21. INFLAMMATORY BOWEL DISEASE
        • Analysis of gene expression profile in patient’s biopsies
    • 22. Inflammatory Bowel Disease
      • Definition:
        • Inflammation of the gastro-intestinal tract that is not due to specific pathogens.
      • Clinical forms:
        • Crohn’s Disease: can affect any part of the digestive tract.
        • Ulcerative Colitis: limited to lower part of large intestine.
      • Environmental factors:
        • Disease of industrialized countries.
        • Smoking reduces risk of UC but increases risks of CD.
      • Treatments:
        • Symptom control.
    • 23. Basic Differences Between CD and UC Ulcerative Colitis Crohn’s Disease Lesion characteristics : - Strictly mucosal. - Continuous and diffuse. - Progress from distal portion of GI tract. Lesion characteristics : - Transmural ( may involve layers as deep as adventitia ). - Scattered and focal. - May appear anywhere along lower GI tract.
    • 24. Study Design
      • Patients
        • Pediatric patients (LSU)
        • 8 Controls, 8 Crohn’s disease patients, 3 colitis patients
      • Tissue samples
        • ~ 500 – 750 mg / biopsy (estimated)
        • Snap frozen upon collection
      • Sample analysis
        • Ratiometric RNA microarrays (2 channels)
        • All samples compared to a common reference sample (total colon) processed in parallel.
    • 25. Legend Apoptosis IFN-  pathway Chemokines Cytokines Tissue remodeling
    • 26. Summary of Observations Tissue repair (Nidogen, IGFBP-5) Proteases (MMP-12, Serpin) Tissue Remodeling Cellular Recruitment MCP-1, MIP3-  Inflammation IFN-  , IL-1  Caspases Apoptosis Inhibitors (Ets-2, BIRC3) IBD Pathology Collaborate with clinical research to obtain critical resources for discovery research
    • 27. Visilizumab in ulcerative colitis
      • Humanized anti-Human CD3 mAb
      • Pharmacology in Steroid Refractory UC patients
        • Rapid and sustained clinical response
        • Rapid and reversible decrease of T cells
      • In vitro mode of action
        • Does not bind to Fc Receptor (engineered Fc portion: IgG2M3)
        • Weak induction of proliferation and cytokine production in PBMC
        • Rapid induction of apoptosis of activated T cells but not of resting T cells
      What is the mechanism of rapid decrease in peripheral T cells in vivo?
    • 28. INFLAMMATORY BOWEL DISEASE
        • Identification of a clinical biomarker from mode of action studies of visilizumab (humanized anti-CD3 mAb)
    • 29. In vitro mode of action: experiment design OKT3: anti-CD3 mAb that binds to FcR  strong activator of T cell proliferation and cytokine release Microarray
    • 30. Results: Cytokines Time RNA Levels (Arbitrary Units)
    • 31. Results: CXC Chemokines Time RNA Levels (Arbitrary Units)
    • 32. Observations and Hypothesis
      • Visilizumab is a weak inducer of cytokine production by T cells
      • Visilizumab is a strong inducer of CXCR3 ligands (IP10, MIG, I-TACK) production by T cells
      • Is this in vitro observation can be confirmed in patients?
      • Is there any association between CXC chemokine production and clinical outcome?
    • 33. Study Design
      • Patients: Steroid refractory UC
        • Unresponsive to IV steroids (minimum 5 days of 40 to 60 mg IV)
        • Composite score Modified Truelove and Witts’ Severity Index (MTWSI) was ≥11.
      • Treatment:
        • Bolus infusions of visilizumab on days 1 and 2 at either 5, 7.5, 10 or 12.5 μg/kg.
      • End Points:
        • Clinical response
        • Peripheral lymphocyte counts
        • Serum IP-10 levels
      Woo J. et al., DDW 2006
    • 34. Rapid increase in serum IP-10 levels in patients treated with visilizumab Woo J. et al., DDW 2006
    • 35. Conclusions
      • Strong correlation between the increase in serum IP-10 levels and reduction in circulating T cells
      • Significant association between elevated serum IP-10 levels and improved clinical outcome  new working model
      Active disease Disease “reset” Maintain a two-way flow of information between clinical and discovery research
    • 36. ASTHMA
        • Analysis of anti-CD25 mAb mode of action in vitro: from nonclinical research to expanded clinical indication
    • 37. Immunopathology of Asthma Sornasse T. et al., AAAAI 2005
      • Allergic component
      • IgE-mediated activation
      • Mast cells and eosinophils
      Metaplasia Airway remodeling
    • 38. Daclizumab
      • Humanized anti-Human CD25 mAb
      • Clinical indication
        • Prophylaxis of acute organ rejection in patients receiving renal transplant
        • Initial positive phase II results in chronic asthma
      • In vitro mode of action: Inhibition of T cell immune functions
        • Inhibition of IL-2 signaling by blocking the binding of IL-2 to the high affinity IL-2 receptor (IL-2 R  )
        • Inhibition of IL-2 dependent T-cell proliferation
    • 39. Effect of daclizumab in vitro on the production of cytokines associated with the immunopathology of asthma
      • System
        • Peripheral Blood Mononuclear Cells from healthy volunteers
      • Activation
        • Anti-CD3 / anti-CD28 coated beads  APC independent
      • Test article
        • Daclizumab: 20 to 0.01 µg/mL
      Sornasse T. et al., AAAAI 2005
    • 40. Daclizumab does not significantly affect  CD3/  CD28-induced proliferation of human PBMC The PBMC of adult volunteers were stimulated with  CD3/  CD28 beads for 72 hours in the presence of graded doses of daclizumab. Results are presented as the Average of the relative proliferation to no-daclizumab control ± SEM Sornasse T. et al., AAAAI 2005 Potential effect on cytokine production is not affected by T cell number
    • 41. Daclizumab strongly inhibits the secretion of pro-asthmatic Th2 cytokines The PBMC of healthy adult volunteers were stimulated with  CD3/  CD28 beads for 72 hours in the presence of graded doses of daclizumab. Results are presented as the Average of the relative cytokine levels to no-daclizumab control ± SEM Sornasse T. et al., AAAAI 2005
    • 42. Daclizumab inhibits the secretion of pro-asthmatic Th0 cytokines The PBMC of healthy adult volunteers were stimulated with  CD3/  CD28 beads for 72 hours in the presence of graded doses of daclizumab. Results are presented as the Average of the relative cytokine levels to no-daclizumab control ± SEM Sornasse T. et al., AAAAI 2005
    • 43. Daclizumab partially inhibits the secretion of pro-inflammatory Th1 cytokines The PBMC of healthy adult volunteers were stimulated with  CD3/  CD28 beads for 72 hours in the presence of graded doses of daclizumab. Results are presented as the Average of the relative cytokine levels to no-daclizumab control ± SEM Sornasse T. et al., AAAAI 2005
    • 44. Conclusions
      • Effect of daclizumab on cytokine production by T cells supports:
        • Pursuing clinical research in allergic asthma
        • Expanding clinical research to non-allergic asthma
      Sornasse T. et al., AAAAI 2005 Discovery and nonclinical research remain essential to articulate a coherent clinical research strategy
    • 45. Building Bridges Between Discovery, Preclinical, and Clinical Research
      • Include nonclinical exploratory end points to clinical studies
      • Collaborate with clinical research to obtain critical resources for discovery research
      • Challenge/verify original hypothesis throughout development
      • Remain aware of model limitation and built-in assumptions
      • Maintain a two-way flow of information between clinical and discovery research
      • Discovery and nonclinical research remain essential to articulate a coherent clinical research strategy
    • 46. Flow of information in research The old view
    • 47. Flow of information in research The new view

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