Limits...
The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma.

Pérez-Mancera PA, Rust AG, van der Weyden L, Kristiansen G, Li A, Sarver AL, Silverstein KA, Grützmann R, Aust D, Rümmele P, Knösel T, Herd C, Stemple DL, Kettleborough R, Brosnan JA, Li A, Morgan R, Knight S, Yu J, Stegeman S, Collier LS, ten Hoeve JJ, de Ridder J, Klein AP, Goggins M, Hruban RH, Chang DK, Biankin AV, Grimmond SM, Australian Pancreatic Cancer Genome InitiativeWessels LF, Wood SA, Iacobuzio-Donahue CA, Pilarsky C, Largaespada DA, Adams DJ, Tuveson DA - Nature (2012)

Bottom Line: Although previous work had attributed a pro-survival role to USP9X in human neoplasia, we found instead that loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis.Clinically, low USP9X protein and messenger RNA expression in PDA correlates with poor survival after surgery, and USP9X levels are inversely associated with metastatic burden in advanced disease.Furthermore, chromatin modulation with trichostatin A or 5-aza-2'-deoxycytidine elevates USP9X expression in human PDA cell lines, indicating a clinical approach for certain patients.

View Article: PubMed Central - PubMed

Affiliation: Li Ka Shing Centre, Cambridge Research Institute, Cancer Research UK, Cambridge CB2 0RE, UK.

ABSTRACT
Pancreatic ductal adenocarcinoma (PDA) remains a lethal malignancy despite much progress concerning its molecular characterization. PDA tumours harbour four signature somatic mutations in addition to numerous lower frequency genetic events of uncertain significance. Here we use Sleeping Beauty (SB) transposon-mediated insertional mutagenesis in a mouse model of pancreatic ductal preneoplasia to identify genes that cooperate with oncogenic Kras(G12D) to accelerate tumorigenesis and promote progression. Our screen revealed new candidate genes for PDA and confirmed the importance of many genes and pathways previously implicated in human PDA. The most commonly mutated gene was the X-linked deubiquitinase Usp9x, which was inactivated in over 50% of the tumours. Although previous work had attributed a pro-survival role to USP9X in human neoplasia, we found instead that loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis. Clinically, low USP9X protein and messenger RNA expression in PDA correlates with poor survival after surgery, and USP9X levels are inversely associated with metastatic burden in advanced disease. Furthermore, chromatin modulation with trichostatin A or 5-aza-2'-deoxycytidine elevates USP9X expression in human PDA cell lines, indicating a clinical approach for certain patients. The conditional deletion of Usp9x cooperated with Kras(G12D) to accelerate pancreatic tumorigenesis in mice, validating their genetic interaction. We propose that USP9X is a major tumour suppressor gene with prognostic and therapeutic relevance in PDA.

Show MeSH

Related in: MedlinePlus

USP9X loss promotes PDAa–c, Decreased USP9X expression correlates with shortened survival in Australian post-surgical cohort (a) (8.7 vs. 18.4 months, p=0.0076; log-rank test), increased metastatic burden in American autopsy series (b) (54% vs. 19%, p=0.0212; Fisher’s exact test), and diminished survival in German post-surgical cohort (c) (11.1 vs. 16.1 months, p=0.037; log-rank test). d, Trichostatin A (TSA, Red) and 5-Aza-2′-deoxycytidine (AZA, Blue) modestly increase USP9X mRNA expression. The mean and s.e.m. of one representative experiment performed in triplicate are shown. e,Usp9x deletion promotes mPanIN progression in KCU mice (p<0.0001; Fisher’s exact test). f, Representative normal pancreas (CU), mPanIN1 (KC), mPanIN3 (KCU) and microinvasive mPDA (KCU, arrow, circled). Scale bar: 100μm.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3376394&req=5

Figure 3: USP9X loss promotes PDAa–c, Decreased USP9X expression correlates with shortened survival in Australian post-surgical cohort (a) (8.7 vs. 18.4 months, p=0.0076; log-rank test), increased metastatic burden in American autopsy series (b) (54% vs. 19%, p=0.0212; Fisher’s exact test), and diminished survival in German post-surgical cohort (c) (11.1 vs. 16.1 months, p=0.037; log-rank test). d, Trichostatin A (TSA, Red) and 5-Aza-2′-deoxycytidine (AZA, Blue) modestly increase USP9X mRNA expression. The mean and s.e.m. of one representative experiment performed in triplicate are shown. e,Usp9x deletion promotes mPanIN progression in KCU mice (p<0.0001; Fisher’s exact test). f, Representative normal pancreas (CU), mPanIN1 (KC), mPanIN3 (KCU) and microinvasive mPDA (KCU, arrow, circled). Scale bar: 100μm.

Mentions: To determine whether USP9X expression is aberrant in human PDA, three distinct patient cohorts were assessed. First, we analyzed a cohort of 100 Australian patients who underwent surgery for localized PDA and had detailed information available concerning clinical-pathological characteristics and outcome (Supplementary Fig. 9, Supplementary Tables 5, 6). Tumor DNA from 88 patients in this cohort failed to yield somatic mutations in USP9X, consistent with prior reports5 (data not shown). Importantly, the low expression of USP9X mRNA correlated with poor survival following surgery (p=0.0076) (Fig. 3a), and multivariate analysis revealed that USP9X expression was an independent poor prognostic factor following surgery (Supplementary Table 7). We next analyzed autopsy specimens from a separate cohort of 42 American patients to determine that USP9X protein expression inversely correlated with a widespread metastatic pattern (p=0.0212) (Fig. 3b), and bore no relation to SMAD4 expression (Supplementary Table 8). A third collection of PDA specimens obtained from resected German patients (n=404) were used to determine that USP9X and ITCH protein levels were decreased (Supplementary Fig. 10a, b) and concordant (Spearman-Rho correlation: 0.47; p<0.01) (Supplementary Table 9a) in tumors compared to normal pancreatic tissue. Additionally, the proportion of tumors that had undetectable USP9X (13.6%) or ITCH (30.5%) protein correlated with a worse outcome (Supplementary Fig. 11, Supplementary Table 9b, c), particularly regarding USP9X in the subset of high grade tumors (Fig. 3c, Supplementary Tables 10 and 11). Collectively, these findings implicate the loss of USP9X expression as a relevant event in human pancreatic cancer progression.


The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma.

Pérez-Mancera PA, Rust AG, van der Weyden L, Kristiansen G, Li A, Sarver AL, Silverstein KA, Grützmann R, Aust D, Rümmele P, Knösel T, Herd C, Stemple DL, Kettleborough R, Brosnan JA, Li A, Morgan R, Knight S, Yu J, Stegeman S, Collier LS, ten Hoeve JJ, de Ridder J, Klein AP, Goggins M, Hruban RH, Chang DK, Biankin AV, Grimmond SM, Australian Pancreatic Cancer Genome InitiativeWessels LF, Wood SA, Iacobuzio-Donahue CA, Pilarsky C, Largaespada DA, Adams DJ, Tuveson DA - Nature (2012)

USP9X loss promotes PDAa–c, Decreased USP9X expression correlates with shortened survival in Australian post-surgical cohort (a) (8.7 vs. 18.4 months, p=0.0076; log-rank test), increased metastatic burden in American autopsy series (b) (54% vs. 19%, p=0.0212; Fisher’s exact test), and diminished survival in German post-surgical cohort (c) (11.1 vs. 16.1 months, p=0.037; log-rank test). d, Trichostatin A (TSA, Red) and 5-Aza-2′-deoxycytidine (AZA, Blue) modestly increase USP9X mRNA expression. The mean and s.e.m. of one representative experiment performed in triplicate are shown. e,Usp9x deletion promotes mPanIN progression in KCU mice (p<0.0001; Fisher’s exact test). f, Representative normal pancreas (CU), mPanIN1 (KC), mPanIN3 (KCU) and microinvasive mPDA (KCU, arrow, circled). Scale bar: 100μm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3376394&req=5

Figure 3: USP9X loss promotes PDAa–c, Decreased USP9X expression correlates with shortened survival in Australian post-surgical cohort (a) (8.7 vs. 18.4 months, p=0.0076; log-rank test), increased metastatic burden in American autopsy series (b) (54% vs. 19%, p=0.0212; Fisher’s exact test), and diminished survival in German post-surgical cohort (c) (11.1 vs. 16.1 months, p=0.037; log-rank test). d, Trichostatin A (TSA, Red) and 5-Aza-2′-deoxycytidine (AZA, Blue) modestly increase USP9X mRNA expression. The mean and s.e.m. of one representative experiment performed in triplicate are shown. e,Usp9x deletion promotes mPanIN progression in KCU mice (p<0.0001; Fisher’s exact test). f, Representative normal pancreas (CU), mPanIN1 (KC), mPanIN3 (KCU) and microinvasive mPDA (KCU, arrow, circled). Scale bar: 100μm.
Mentions: To determine whether USP9X expression is aberrant in human PDA, three distinct patient cohorts were assessed. First, we analyzed a cohort of 100 Australian patients who underwent surgery for localized PDA and had detailed information available concerning clinical-pathological characteristics and outcome (Supplementary Fig. 9, Supplementary Tables 5, 6). Tumor DNA from 88 patients in this cohort failed to yield somatic mutations in USP9X, consistent with prior reports5 (data not shown). Importantly, the low expression of USP9X mRNA correlated with poor survival following surgery (p=0.0076) (Fig. 3a), and multivariate analysis revealed that USP9X expression was an independent poor prognostic factor following surgery (Supplementary Table 7). We next analyzed autopsy specimens from a separate cohort of 42 American patients to determine that USP9X protein expression inversely correlated with a widespread metastatic pattern (p=0.0212) (Fig. 3b), and bore no relation to SMAD4 expression (Supplementary Table 8). A third collection of PDA specimens obtained from resected German patients (n=404) were used to determine that USP9X and ITCH protein levels were decreased (Supplementary Fig. 10a, b) and concordant (Spearman-Rho correlation: 0.47; p<0.01) (Supplementary Table 9a) in tumors compared to normal pancreatic tissue. Additionally, the proportion of tumors that had undetectable USP9X (13.6%) or ITCH (30.5%) protein correlated with a worse outcome (Supplementary Fig. 11, Supplementary Table 9b, c), particularly regarding USP9X in the subset of high grade tumors (Fig. 3c, Supplementary Tables 10 and 11). Collectively, these findings implicate the loss of USP9X expression as a relevant event in human pancreatic cancer progression.

Bottom Line: Although previous work had attributed a pro-survival role to USP9X in human neoplasia, we found instead that loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis.Clinically, low USP9X protein and messenger RNA expression in PDA correlates with poor survival after surgery, and USP9X levels are inversely associated with metastatic burden in advanced disease.Furthermore, chromatin modulation with trichostatin A or 5-aza-2'-deoxycytidine elevates USP9X expression in human PDA cell lines, indicating a clinical approach for certain patients.

View Article: PubMed Central - PubMed

Affiliation: Li Ka Shing Centre, Cambridge Research Institute, Cancer Research UK, Cambridge CB2 0RE, UK.

ABSTRACT
Pancreatic ductal adenocarcinoma (PDA) remains a lethal malignancy despite much progress concerning its molecular characterization. PDA tumours harbour four signature somatic mutations in addition to numerous lower frequency genetic events of uncertain significance. Here we use Sleeping Beauty (SB) transposon-mediated insertional mutagenesis in a mouse model of pancreatic ductal preneoplasia to identify genes that cooperate with oncogenic Kras(G12D) to accelerate tumorigenesis and promote progression. Our screen revealed new candidate genes for PDA and confirmed the importance of many genes and pathways previously implicated in human PDA. The most commonly mutated gene was the X-linked deubiquitinase Usp9x, which was inactivated in over 50% of the tumours. Although previous work had attributed a pro-survival role to USP9X in human neoplasia, we found instead that loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis. Clinically, low USP9X protein and messenger RNA expression in PDA correlates with poor survival after surgery, and USP9X levels are inversely associated with metastatic burden in advanced disease. Furthermore, chromatin modulation with trichostatin A or 5-aza-2'-deoxycytidine elevates USP9X expression in human PDA cell lines, indicating a clinical approach for certain patients. The conditional deletion of Usp9x cooperated with Kras(G12D) to accelerate pancreatic tumorigenesis in mice, validating their genetic interaction. We propose that USP9X is a major tumour suppressor gene with prognostic and therapeutic relevance in PDA.

Show MeSH
Related in: MedlinePlus