Understanding the mechanisms of asbestos related diseases

Transcription

1 University of Hawai i Cancer Center Understanding the mechanisms of asbestos related diseases Haining Yang, PhD Professor University of Hawai i Cancer Center

2 Marker of exposure: Bilateral pleural plaques Non-malignant diseases: i. Asbestosis (diffuse bilateral lung fibrosis, can lead to death) ii. Diffuse pleural thickening iii. Pleural effusion iv. Rounded atelectasis (folded lung) Malignant diseases i. Malignant Mesothelioma (MM) ii. Lung Cancer

3 Potential targets and strategies for mesothelioma therapy that have been proposed based on recent studies. Michele Carbone, and Haining Yang Clin Cancer Res 2012;18: by American Association for Cancer Research

4 Normal human mesothelial cells (HM) HM Exposed to Crocidolite Asbestos

5 Asbestos is cytotoxic to HM in tissue culture. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

6 TNF-α and TNF-α receptor TNF-R1 are induced in HM following asbestos exposure. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

7 TNF-α significantly reduces asbestos cytotoxicity. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

8 TNF-α induces the activation of NF-κB. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

9 Inhibition of the NF-κB pathway by Bay suppresses the anticytotoxicity effect induced by TNF-α. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

10 RNA interference assays confirm that TNF-α inhibits asbestos induced cytotoxicity by means of a NF-κB dependent mechanism. Haining Yang et al. PNAS 2006;103: by National Academy of Sciences

11 crocidolite+tnf- crocidolite+tnf- crocidolite asbestos 1 week 1 month

12 The American Journal of Pathology , DOI: ( /j.ajpath ) Copyright 2013 American Society for Investigative Pathology Terms and Conditions

13 The American Journal of Pathology , DOI: ( /j.ajpath ) Copyright 2013 American Society for Investigative Pathology Terms and Conditions

14 The American Journal of Pathology , DOI: ( /j.ajpath ) Copyright 2013 American Society for Investigative Pathology Terms and Conditions

15 How does asbestos kill mesothelial cells? What kind of Cell death? Apoptosis? Necrosis?

16 Asbestos induced cell death -involves programmed necrosis New concept of necrosis -- It is a regulated event The process of necrosis normally involves-- 1) Activation of PARP (poly(adpribose)polymerase) 2) ATP depletion 3) Release of HMGB1(high mobility group box 1) 4) Induction of inflammation

17 Asbestos induces PARP activation in HM. (A) Increasing amounts of asbestos induced a parallel increase in PARP levels, without inducing PARP cleavage. Haining Yang et al. PNAS 2010;107: by National Academy of Sciences

18 Asbestos-induced HM death is PARP dependent and causes ATP depletion. Haining Yang et al. PNAS 2010;107: by National Academy of Sciences

19 HMGB1

20 HMGB1 is released from HM into the extracellular space after asbestos exposure. Haining Yang et al. PNAS 2010;107: by National Academy of Sciences

21 Asbestos-induced cell death and the release of HMGB1 promote TNF-α secretion by macrophages. Haining Yang et al. PNAS 2010;107: by National Academy of Sciences

22 Yang H et al, PNAS 2006, PNAS 2010, Clinical Cancer Res 2011

23 Asbestos Inhibition of HMGB1 decreases the foci formation in the in vitro MM model (HM/macrophages co-culture system)

24 Immunohistochemical analyses show strong HMGB1 and TNF-α staining around areas of asbestos deposits in two representative murine specimens. Haining Yang et al. PNAS 2010;107: by National Academy of Sciences

25 HMGB1 levels in the serum of mice injected with different types of fibers 40 HMGB1 ELISA - Mice injected w/ fibers 35 * HMGB1 (ng/ml) * * * * * 5 0 PBS Glass CHRY CRO ND OR TK TRE

26 HMGB1 serum levels are long-term persistent after exposure to crocidolite but not to chrysotile (Qi et al, Am J Pathol, 2013) HMGB1 levels in the serum of mice injected with with a total of 5 mg of crocidolite or chrysotile in a high-dose, short-term protocol with two weekly injections of 2.5 mg each

27 HMGB1 levels in the serum of mice injected with with a total of 5 mg of crocidolite or chrysotile in a low-dose, long-term protocol with 10 weekly injections of 0.5 mg each

28 Model for asbestos carcinogenesis: Yang H et al, PNAS 2006, 2010, Clinical Cancer Res 2011

29 Yang H et al, PNAS 2010

30 HMGB1 is highly expressed in mesothelioma tissue. Normal pleura MM 100X 400X

31 MM cells contain high levels of HMGB1 in both nucleus and cytoplasm MM cell lines secrete HMGB1 HMGB1 ELISA using cell culture medium

32 MM cells migration assay

33 Inhibition of HMGB1 reduces MM cells proliferation and mobility Wound-healing assay

34 Inhibition of HMGB1 reduces anchorage independent growth of MM cells

35 Anti-HMGB1 mab inhibits MM growth in mouse model

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37 Jube S, M Carbone and H Yang. Cancer Res. 2012

38 Anti-inflammatory drug: Aspirin low toxicity, and proven efficacy in reducing the incidence of colorectal cancer. The Physician's Health Study In the study, 22,067 physicians were followed for 24 years. <aspirin (325 mg on alternate days) or placebo> 17 cases of MM reported. An intention to treat analysis comparing aspirin to placebo revealed a relative risk of 0.7 ( % confidence interval). The risk was 30% lower in the aspirin treatment group The result indicates a possible association between aspirin use and reduced MM incidence.

39 Effect of different anti-inflammatory drugs on MM motility Aspirin: acetylsalicylic acid (ASA)

40 Anti-inflammatory drug: Aspirin Aspirin reduces HMGB1 serum levels in mice exposed to asbestos Aspirin reduces HMGB1 serum levels in mice with MM

41 Impossibile visualizzare l'immagine.

42 Salicylates inhibit colony formation of MM cells

43 Salicylates inhibit colony formation of high HMGB1-secreting MM cells (A-B) but not of low HMGB1-secreting cells (C-D).

44 Effect of SA on HMGB1-induced migration is COX-2 independent

45 Aspirin inhibits MM tumor growth in mice

46 Conclusion: HMGB1 is a critical player in asbestos carcinogenesis and MM pathogenesis. Targeting HMGB1 can be a novel strategy for mesothelioma prevention and therapy Some of the aspirin anticancer activity may be mediated through inhibition of HMGB1. Our results are relevant to mesothelioma and to other inflammation-related cancers in which HMGB1 plays a role. Specific HMGB1 inhibitors, such as BoxA, may have stronger anti-cancer activity and less side effects than aspirin.

47 HMGB1 serum level is significantly higher in MM patients than in healthy controls.

48 - NCI R01 (Yang PI) Current Active Grants HMGB1 and Its Isoforms As Biomarkers For Mineral Fiber Exposure and MM Detection (Yang and Carbone, Co PIs) Identification and validation of novel germline DNA variants associated to increased risk of malignant mesothelioma - The EDRN Mesothelioma Biomarker Discovery Laboratory - V Foundation Translational Grant (Yang-Basic Science PI, Carbone-Clinical PI) Riviera United-4 A Cure Foundation grant