Paola Zanelli

Novel association of HLA-haplotypes with primary sclerosing cholangitis (PSC) in a southern European population

AIMS In patients with with primary sclerosing cholangitis we investigated the major histocompatibility complex (MHC) genes and mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. METHODS In 64 PSC patients and 183 normal controls of the same population (Northern Italy), allelic polymorphisms at the DNA level were investigated in MHC region genes: HLA-DRB1, HLA-DQB1 and HLA-B, tumour necrosis factor A (TNFA), and in CFTR gene, with polymerase chain reaction-based methodologies. RESULTS Frequencies of DRB1*01, DQA1*0101, DQB1*0102 (14 vs. 8%, p<0.05), DRB1*16, DQA1*0102, DQB1*0502 (8 vs. 3%, p<0.025) and DRB1*04, DQA1*03, DQB1*0301 (10 vs. 4%, p<0.005) haplotypes were more elevated in PSC patients. The frequency of patients positive for HLA DRB1*01, *1601 or *04 related haplotypes was significantly increased (32 vs. 14%, p<0.00025). DRB1*07, DQA1*0201, DQB1*02 haplotype frequency was significantly decreased (4 vs. 15%, p<0.001). After removing HLA-DRB1*01, *1601, *04 related haplotype sharing patients, HLA-DRB1*03, DQA1*0501, DQB1*02 haplotype frequency was significantly increased (32 vs. 14%, p<0.01). TNFA2 allele frequency was significantly increased in PSC patients (23 vs. 14%, p<0.025), as well as the TNFA2 homozygous genotype (9 vs. 0.5%, p=0.0013). No mutations were found on the CFTR gene and the allelic frequency of the 5T polymorphism in intron 8 was not increased. CONCLUSION These data suggest that the role of genes in the HLA region is relevant, but not necessarily disease-specific and it might be different in populations with divergent ancestries.

HLA and killer immunoglobulin-like receptor genes as outcome predictors of hepatitis C virus-related hepatocellular carcinoma.

Purpose: We evaluated the impact of the killer immunoglobulin-like receptors (KIR) of natural killer (NK) cells and of their HLA ligands over the clinical outcome of hepatitis C virus (HCV)–related hepatocellular carcinoma after curative treatment by either surgical resection or radiofrequency thermal ablation (RTA). Experimental Design: Sixty-one consecutive patients with HCV-related hepatocellular carcinoma underwent KIR genotyping and HLA typing. A phenotypic/functional characterization of NK cells was carried out in patients with different KIR/KIR-ligand genotype. Results: Activating KIR2DS5 was associated with significantly longer time to recurrence (TTR) and overall survival (OS; P < 0.03 each). Homozygous HLA-C1 (P < 0.02) and HLA-Bw4I80 (P < 0.05) were expressed by patients with significantly better OS, whereas HLA-C2 (P < 0.02) and HLA-Bw4T80 (P < 0.01) were associated with a worse OS. Multivariate analysis identified as parameters independently related to TTR the type of treatment (surgical resection vs. RTA; P < 0.03) and HLA-C1 (P < 0.03), whereas only KIR2DS5 was an independent predictor of longer OS (P < 0.05). Compound KIR2DL2-C1 and KIR3DS1-Bw4T80 genotypes were associated with better TTR (P < 0.03) and worse OS (P = 0.02), respectively. A prevalent cytotoxic (CD56dim) NK phenotype was detected in patients with both longer TTR and OS. Cytotoxic capacity measured by upregulation of CD107a was significantly higher in subjects with HLA-C1 alone or combined with KIR2DL2/KIR2DL3. Conclusions: These results support a central role of NK cells in the immune response against hepatocellular carcinoma, providing a strong rationale for therapeutic strategies enhancing NK response and for individualized posttreatment monitoring schemes. Clin Cancer Res; 19(19); 5465–73. ©2013 AACR.

A breast cancer patient from Italy with germline mutations in both the BRCA1 and BRCA2 genes

We report the first case in Italy of a non-Ashkenazi double heterozygote for BRCA1 and BRCA2 genes. This finding is predictably rare, with a maximum frequency of 1/250,000. The proband and her mother were diagnosed with early-onset breast cancer. No other relatives with breast and/or ovarian cancer were observed. The implications of this case in regard to genetic testing and counseling are substantial.

BRCA1 status, molecular markers, clinical variables in breast cancer patients with high probability of having an inherited genetic mutation.

9648 Background: to evaluate the clinical features and outcomes of Breast Cancer (BC) patients with genetic susceptibility to this disease and to investigate the contribution of BRCA1 germline mutation to the phenotype of these tumors. METHODS we reviewed clinical and pathological records of 144 women with autosomal dominant inheritance of breast (+/- ovarian) cancer risk. All women underwent full genetic counseling. Of these, 101 selected patients with high probability of having a germ-line mutation were tested for BRCA1 mutation analysis. Exon 11 was screened for BRCA1 mutations using Protein Truncation Test; mutations detected were confirmed by Direct Sequencing. All the other exons were analyzed by DS. RESULTS the two different risk groups had similar clinical outcomes. Of the 57 patients with completed mutation analysis, 44 (77%) patients had wild type BRCA1, 8 (14%) had variants of unclear significance, 5 (8%) had deleterious mutations in BRCA1. With regard to entry criteria for BRCA1 genetic testing, mutations were detected in 5% (1/20), 2,5% (1/41), 16% (2/12) and 16% (1/6) of women with family history, early onset BC (< 40 years), Breast-Ovarian Cancer (BOC) and early onset plus Bilateral Breast Cancer, respectively. BRCA1 Associated Breast Cancers (BABC) were more likely to have histological grade 3 and high proliferation rate than negative cases (40% v 27%; 60% v 45%). These differences were not statistically significant. BABC were significantly more likely to be estrogen receptor-negative (67% v 16%, P = .04). Though not significant, all valuable tumors with BRCA1 mutations were HER-2/neu negative. In the entire cohort, there were no significant differences between BABC and non-BABC in 5 year relapse free survival (60% v 78%, P = not significant [NS]), 5 year event free survival (60% v 66%, P = NS), or 5 year overall survival. CONCLUSIONS BABC seem to present with adverse molecular and histopathologic features when compared with cases not associated with BRCA1 mutations. However, the prognosis of BABC appears to be similar to that of non associated cancer. No significant financial relationships to disclose.