News dalla ricerca: identificazione di nuove molecole come target di terapia nelle infiammazioni intestinali

Transcription

1 News dalla ricerca: identificazione di nuove molecole come target di terapia nelle infiammazioni intestinali Salvatore Cucchiara, Sapienza Università di Roma Dipartimento di Pediatria e Neuropsichiatria Infantile Unità Complessa di Gastroenterologia e Epatologia Pediatrica

2 BIOLOGICAL THERAPY FOR INFLAMMATORY BOWEL DISEASE: REMARKS ü highlighting the central role of TNFα in the pathogenesis of IBD, inhibition of its signaling was the first successful biological treatment for IBD

3 Three classes of anti-tnf: fusion protein, antibodies and PEGylated Fab fragment

4 POTENTIAL BIOLOGICAL THERAPEUTIC TARGETS FOR IBD host-microbial interactions at the luminal-epithelial cell interface antigen processing by macrophages and presentation to T-cells T-cell activation/signaling T-cell differentiation cytokine production leukocyte trafficking

5 SELECTIVE ADHESION MOLECULE INHIBITION: IMPLICATIONS FOR CROHN S DISEASE THERAPY mucosal and inflammatory zip codes

6 LEUKOCYTE ADHESION IN HIGH ENDOTHELIAL VENULES OF THE GUT IS A MULTISTEP PROCESS

7 MLN-02 (LDP-02): Humanized monoclonal antibody (mab) against α4β7 integrins GUT SPECIFICITY

8 NEWER TH1/TH2/TH17 PARADIGM

9 BIOLOGY OF INTERLEUKINS 12 AND 23 USTEKINUMAB ABT-874 IFNγ USTEKINUMAB ABT-874 Ustekinumab: human IgG1k monoclonal antibody, binds to the p40 subunit of human and primate IL-12/23, prevents IL-12 and IL-23 from binding IL-12Rb1

10

11

12 Lotze and Tracey, 2005

13 P. Matzinger, Science 2002 The Danger Model Initiation of the inflammatory response occurs in response to molecular patterns that are associated with both pathogens and some normal cellular components that are released by damaged cells during both infectious and sterile processes Immune activation is the result of recognition of molecular patterns by cellular receptors Pathogen-Associated Molecular Patterns PAMPs LPS Bacterial DNA Viral RNA Flagellin Damage-Associated Molecular Patterns DAMPs HMGB1 HSPs S100s IL-1alpha Pattern Recognition Receptors PRRs Pattern Recognition Receptors PRRs DAMPs comprises intracellular molecules characterized by the ability to reach the extracellular environment, where they prompt inflammation INFLAMMATION

14 Dumitriu et al Monocytes, macrophages, NK cells, dendritic cells, endothelial cells, synovial fibroblasts, platelets, astrocytes and tumor cells The secretion occurs in association with a relocalization of HMGB1 from the nucleus to the cytoplasmic secretory lysosomes Activation of monocytes by LPS, TNFalpha, INFgamma or IL-1 leads to posttraslational modifications (phosphorilation, acetylation, methylation) to block the re-entry into the nuclear compartment Extracellular HMGB1 has proinflammatory properties: proinflammatory cytokine production, up-regulation of adhesion molecules on endothelial cells, activation of innate and immune cells

15 HMGB1 and Immune/Inflammatory Disorders ü The role of HMGB1 in the pathogenesis of rheumatic disease. Andersson and Harris. Biochim Biophys Acta 2010 ü HMGB1 in systemic lupus erythematosus: Its role in cutaneous lesions development. Abdulahad et al. Autoimmun Rev 2010 ü Increase of high-mobility group box chromosomal protein 1 in blood and injured organs in experimental severe acute pancreatitis. Yasuda et al. Pancreas 2007 ü Early release of HMGB-1 from neurons after the onset of brain ischemia. Qiu et al. J Cereb Flow Metab 2008 ü High Mobility Group Box-1 Protein (HMGB1) is increased in ANCA-associated vasculitis (AAV) with renal manifestations. Bruchfeld et al. Mol Med 2010 ü Cytokine profiles of bronchoalveolar lavage fluid in patients with pneumocystis pneumonia. Tasaka et al. Microbiol Immunol 2010 ü Proline-Glycine-Proline (PGP) and High Mobility Group Box Protein-1 (HMGB1): Potential Mediators of Cystic Fibrosis Airway Inflammation. Gaggar et al. Open Respir Med J 2010 ü HMGB1, a potent proinflammatory cytokine in sepsis. Huang et al. Cytokine 2010 ü High mobility group box protein-1 in experimental autoimmune uveoretinitis. Watanabe T et al. Invest Ophtalmol Vis Sci 2009 ü Clinical significance of serum HMGB-1 and srage levels in systemic sclerosis: association with disease severity. Yoshizaki et al. J Clin Immunol 2009 ü HMGB1, an innate alarmin, in the pathogenesis of type 1 diabetes. Zhang et al. Int J Clin Exp Pathol 2009

16 H. Yang and K. Tracey, 2010

17 HMGB1 and Inflammatory Bowel Disease AIM: assessing the presence of HMGB1 in the stools of patients with IBD in order to evaluate its role as non invasive subclinical marker of intestinal inflammation Pa#ents: 20 pediatric pa-ents with CD 21 with UC and 13 controls referred to the Pediatric Gastroenterology and Liver Unit of the Sapienza University of Rome, formed the basis of this study.

18 Table1: Demographic and Clinical Characteris#cs of the IBD Study Popula#on Crohn s disease (CD) Subject Sex Age PCDAI Location Severe CD1 F L3B1 CD2 M L3B2 CD3 M L3B1 CD4 M L3B1 CD5 F L2L4B1 CD6 M L2B1 CD7 F L3B3p CD8 F L3B3p Mild/Moderate CD9 M CD10 M 9 17 L3B2 CD11 M L3L4B1 Inactive CD12 F P CD13 F L1B2 CD14 M L2B1 CD15 M L1B1 CD16 M L3B2 CD17 M 18 5 L1L4B2 CD18 M 16 5 L3B1 CD19 M 12 5 L3 PCDAI: pediatric Crohn s disease ac-vity index Ulcera#ve Coli#s (UC) Subject Sex Age PUCAI Location Severe UC1 F E2 UC2 F E3 UC3 M E3 UC4 F 7 65 E2 UC5 M E3 Mild/Moderate UC6 F E1 UC7 M E3 UC8 M E2 UC9 M E2 UC10 F E3 UC11 F E3 UC12 M E1 UC13 M E1 Inactive UC14 F E2 UC15 F 18 5 E2 UC16 F 11 5 E3 UC17 M 12 0 E3 UC18 M 10 0 E2 UC19 F 10 0 E3 UC20 F 7 0 E1 UC21 M 14 0 E2 PUCAI: pediatric Ulcera-ve Coli-s ac-vity index Montreal Classifica#on for Crohn s disease: Loca#on:L1: ileal; L2: colonic; L3: ileocolonic; L4: isolated upper disease (L4 is a modifier that can be added to L1- L3 when concomitant upper gastrointes-nal disease is present). Behaviour: B1: non- stricturing, non- penetra-ng; B2: structuring; B3: penetra-ng; p: perianal disease modifier (p is added to B1- B3 when concomitant perianal disease is present). Montreal Classifica#on for ulcera#ve coli#s: E1: Ulcera-ve proc--s; E2: LeP- sided UC; E3: Extensive UC.

19 Fecal HMGB1 is a marker of intestinal inflammation and of disease severity

20 Fecal HMGB1 is a marker of intestinal inflammation and of disease severity

21 Fecal HMGB1 is a marker of intestinal inflammation and of disease severity

22 Fecal HMGB1 vs Fecal Calprotectin

23 HMGB1 gene/protein expression levels are not increased in bioptic tissues of IBD patients

24 HMGB1 is shifted from the nucleus to the cytoplasm during inflammation - 1

25 HMGB1 is shifted from the nucleus to the cytoplasm during inflammation - 2 -

26 Extracellular secre#on of HMGB1 is induced in vitro and ex vivo by inflammatory s#muli

27 HMGB1 and Inflammatory Bowel Disease CONCLUSIONS: v fecal HMGB1 can be considered a novel marker of human intes#nal inflamma#on and a useful indicator of the disease status. v extracellular release of HMGB1 is over- induced in human intes#nal cells by inflammatory s#muli v Intes#nal immune cells, when ac#vated, can secrete their nuclear HMGB1, without need for further syntesis, which accumulates in the cytoplasm and then is released in the extracellular matrix where it displays its inflammatory proper#es. v methyla#on, other than acetyla#on and phosphoryla#on, is one of the post- transla#onal forms of the inflammatory HMGB1

28 IN VITRO v To assess the ability of recombinant HMGB1 to induce inflamma#on EX VIVO v To assess the presence of phpshorylated and acetylated HMGB1 in the inflamed intes#nal mucosa of IBD pa#ents v To inves#gate HMGB1 in the stools of a large cohort of adult pa#ents with both CD and UC IN VIVO HMGB1 and Inflammatory Bowel Disease: ongoing work v To test the efficacy of glycirrhizin in decreasing intes#nal inflamma#on in more DSS- induced coli#s mice models (C57BL/6; Balb/c; IL- 10- /- ) v To inves#gate whether glycirrhizin is able to prevent intes#nal inflamma#on

29