Fernando Rivera

Mutations in BRIP1 confer high risk of ovarian cancer

Ovarian cancer causes more deaths than any other gynecologic malignancy in developed countries. Sixteen million sequence variants, identified through whole-genome sequencing of 457 Icelanders, were imputed to 41,675 Icelanders genotyped using SNP chips, as well as to their relatives. Sequence variants were tested for association with ovarian cancer (N of affected individuals = 656). We discovered a rare (0.41% allelic frequency) frameshift mutation, c.2040_2041insTT, in the BRIP1 (FANCJ) gene that confers an increase in ovarian cancer risk (odds ratio (OR) = 8.13, P = 2.8 × 10−14). The mutation was also associated with increased risk of cancer in general and reduced lifespan by 3.6 years. In a Spanish population, another frameshift mutation in BRIP1, c.1702_1703del, was seen in 2 out of 144 subjects with ovarian cancer and 1 out of 1,780 control subjects (P = 0.016). This allele was also associated with breast cancer (seen in 6/927 cases; P = 0.0079). Ovarian tumors from heterozygous carriers of the Icelandic mutation show loss of the wild-type allele, indicating that BRIP1 behaves like a classical tumor suppressor gene in ovarian cancer.

Discovery of common variants associated with low TSH levels and thyroid cancer risk

To search for sequence variants conferring risk of nonmedullary thyroid cancer, we focused our analysis on 22 SNPs with a P < 5 × 10−8 in a genome-wide association study on levels of thyroid stimulating hormone (TSH) in 27,758 Icelanders. Of those, rs965513 has previously been shown to associate with thyroid cancer. The remaining 21 SNPs were genotyped in 561 Icelandic individuals with thyroid cancer (cases) and up to 40,013 controls. Variants suggestively associated with thyroid cancer (P < 0.05) were genotyped in an additional 595 non-Icelandic cases and 2,604 controls. After combining the results, three variants were shown to associate with thyroid cancer: rs966423 on 2q35 (OR = 1.34; Pcombined = 1.3 × 10−9), rs2439302 on 8p12 (OR = 1.36; Pcombined = 2.0 × 10−9) and rs116909374 on 14q13.3 (OR = 2.09; Pcombined = 4.6 × 10−11), a region previously reported to contain an uncorrelated variant conferring risk of thyroid cancer. A strong association (P = 9.1 × 10−91) was observed between rs2439302 on 8p12 and expression of NRG1, which encodes the signaling protein neuregulin 1, in blood.

Genome-wide association study yields variants at 20p12.2 that associate with urinary bladder cancer.

Genome-wide association studies (GWAS) of urinary bladder cancer (UBC) have yielded common variants at 12 loci that associate with risk of the disease. We report here the results of a GWAS of UBC including 1670 UBC cases and 90 180 controls, followed by replication analysis in additional 5266 UBC cases and 10 456 controls. We tested a dataset containing 34.2 million variants, generated by imputation based on whole-genome sequencing of 2230 Icelanders. Several correlated variants at 20p12, represented by rs62185668, show genome-wide significant association with UBC after combining discovery and replication results (OR = 1.19, P = 1.5 × 10(-11) for rs62185668-A, minor allele frequency = 23.6%). The variants are located in a non-coding region approximately 300 kb upstream from the JAG1 gene, an important component of the Notch signaling pathways that may be oncogenic or tumor suppressive in several forms of cancer. Our results add to the growing number of UBC risk variants discovered through GWAS.

A genome-wide association study yields five novel thyroid cancer risk loci

The great majority of thyroid cancers are of the non-medullary type. Here we report findings from a genome-wide association study of non-medullary thyroid cancer, including in total 3,001 patients and 287,550 controls from five study groups of European descent. Our results yield five novel loci (all with Pcombined<3 × 10−8): 1q42.2 (rs12129938 in PCNXL2), 3q26.2 (rs6793295 a missense mutation in LRCC34 near TERC), 5q22.1 (rs73227498 between NREP and EPB41L4A), 10q24.33 (rs7902587 near OBFC1), and two independently associated variants at 15q22.33 (rs2289261 and rs56062135; both in SMAD3). We also confirm recently published association results from a Chinese study of a variant on 5p15.33 (rs2736100 near the TERT gene) and present a stronger association result for a moderately correlated variant (rs10069690; OR=1.20, P=3.2 × 10−7) based on our study of individuals of European ancestry. In combination, these results raise several opportunities for future studies of the pathogenesis of thyroid cancer.

GNP-GAPDH 1-22 nanovaccines prevent neonatal listeriosis by blocking microglial apoptosis and bacterial dissemination

Clinical cases of neonatal listeriosis are associated with brain disease and fetal loss due to complications in early or late pregnancy, which suggests that microglial function is altered. This is believed to be the first study to link microglial apoptosis with neonatal listeriosis and listeriosis-associated brain disease, and to propose a new nanovaccine formulation that reverses all effects of listeriosis and confers Listeria monocytogenes (LM)-specific immunity. We examined clinical cases of neonatal listeriosis in 2013-2015 and defined two useful prognostic immune biomarkers to design listeriosis vaccines: high anti-GAPDH1-22 titres and tumor necrosis factor (TNF)/interleukin (IL)-6 ratios. Therefore, we developed a nanovaccine with gold glyco-nanoparticles conjugated to LM peptide 1-22 of GAPDH (Lmo2459), GNP-GAPDH1-22 nanovaccinesformulated with a pro-inflammatory Toll-like receptor 2/4-targeted adjuvant. Neonates born to non-vaccinated pregnant mice with listeriosis, showed brain and vascular diseases and significant microglial dysfunction by induction of TNF-α-mediated apoptosis. This programmed TNF-mediated suicide explains LM dissemination in brains and livers and blocks production of early pro-inflammatory cytokines such as IL-1β and interferon-α/β. In contrast, neonates born to GNP-GAPDH1-22-vaccinated mothers before LM infection, did not develop listeriosis or brain diseases and had functional microglia. In nanovaccinated mothers, immune responses shifted towards Th1/IL-12 pro-inflammatory cytokine profiles and high production of anti-GAPDH1-22 antibodies, suggesting good induction of LM-specific memory.Clinical cases of neonatal listeriosis are associated with brain disease and fetal loss due to complications in early or late pregnancy, which suggests that microglial function is altered. This is believed to be the first study to link microglial apoptosis with neonatal listeriosis and listeriosis-associated brain disease, and to propose a new nanovaccine formulation that reverses all effects of listeriosis and confers Listeria monocytogenes (LM)-specific immunity. We examined clinical cases of neonatal listeriosis in 2013–2015 and defined two useful prognostic immune biomarkers to design listeriosis vaccines: high anti-GAPDH1-22 titres and tumor necrosis factor (TNF)/interleukin (IL)-6 ratios. Therefore, we developed a nanovaccine with gold glyco-nanoparticles conjugated to LM peptide 1-22 of GAPDH (Lmo2459), GNP-GAPDH1-22 nanovaccinesformulated with a pro-inflammatory Toll-like receptor 2/4-targeted adjuvant. Neonates born to non-vaccinated pregnant mice with listeriosis, showed brain and vascular diseases and significant microglial dysfunction by induction of TNF-α-mediated apoptosis. This programmed TNF-mediated suicide explains LM dissemination in brains and livers and blocks production of early pro-inflammatory cytokines such as IL-1β and interferon-α/β. In contrast, neonates born to GNP-GAPDH1–22-vaccinated mothers before LM infection, did not develop listeriosis or brain diseases and had functional microglia. In nanovaccinated mothers, immune responses shifted towards Th1/IL-12 pro-inflammatory cytokine profiles and high production of anti-GAPDH1–22 antibodies, suggesting good induction of LM-specific memory.

Colony-stimulating factors for adjunctive therapy of infections in neutropenic cancer patients

Dear Editor, We read with interest the article by Schimpf, Scott and Wade concerning controversial issues on infections in cancer patients [7]. One of the issues addressed in this article was the use of cytokines [granulocyte-colony-stimulating factor (GCSF) or granulocyte macrophagecolony-stimulating factor (GMCSF)] for adjunctive therapy of infections in cancer patients with chemotherapy-induced neutropenia and fever, an area in which no randomized trials had been performed until recently. This is somewhat surprising, since both G-CSF and GM-CSF have been shown to reduce the duration of chemotherapy-induced neutropenia [2], which is an important prognostic factor for patients with neutropenic fever [1]. Limited animal studies also support a role for colony-stimulating factors (CSF) in the t reatment of infection in neutropenic hosts [5]. We recently completed a randomized trial to test the addition of G-CSF or GM-CSF to antibiotics in cancer patients on standarddose chemotherapy who developed neutropenic fever [4]. A total of 121 patients were enrolled and randomized to G-CSF (5 txg kg -1 day -1 s.c.), GM-CSF (same dose) or placebo, in addition to standard i.v. antibiotics ( recommended schedule = ceftazidime + amikacin). CSF or the placebo was to be discontinued when the absolute neutrophil count (ANC) was above 1000/mm 3 for 2 consecutive days (whether the patient remained febrile or not). Intravenous antibiotics were to be discontinued and the patient was to be discharged when he or she had been afebrile and with A N C above 1000/mm 3 for 2 consecutive days, provided that 5 days on i.v. antibiotics had been completed (7 days if cultures had been positive or a site of infection was present). Results are summarized in Table 1. Hematological recovery was faster in patients on G-CSF or GM-CSF (median time to A N C > 1000/mm3=3 days for GCSF compared to 2 for GM-CSF and 5 for placebo, P < 0.001 versus each CSF arm). The number of patients with prolonged neutropenia ( A N C < 1000/mm 3 for more than 10 days) was significantly larger in the placebo group [9 patients (20.9%) for placebo compared to 0 for G-CSF and GM-CSF, P<0.001]. The duration of fever was short and there were no significant differences among the three arms. The median hospital stay (and duration of i.v. antibiotic therapy) was significantly shorter in each of the groups t reated with CSF when compared with placebo (5 days for G-CSF and for GMCSF versus 7 for placebo, P < 0.001), and this translated into a decreased cost per patient (mean = U.S.

RAS mutations distribution in a 108 patients series of colorectal cancer

5600 for G-CSF compared to U.S.

Re: Decision Analysis of Hematopoietic Growth Factor Use in Patients Receiving Cancer Chemotherapy

5500 for GMCSF and U.S.

Gold glyconanoparticles coupled to listeriolysin O 91–99 peptide serve as adjuvant therapy against melanoma

6900 for placebo), bearing in mind that the time on CSF was short (median = 4 days both for G-CSF and for GM-CSF). We could not find any difference in efficacy or toxicity between GCSF and GM-CSF, except for CSFrelated fever, arbitrarily defined as fever subsiding less than 24 h after the last dose of CSF, which was found in 4 patients (10%) on GMCSF and 1 (2%) on G-CSF. Since CSF was discontinued upon hematological recovery, whether the patient remained febrile or not, CSFrelated fever did not result in prolonged duration of antibiotic thera-

COMMENT ON : DECISION ANALYSIS OF HEMATOPOIETIC GROWTH FACTOR USE IN PATIENTS RECEIVING CANCER CHEMOTHERAPY . AUTHORS' REPLY

Background: As described previously in high impact studies,1 patients with other activating RAS out of codon 2 of kras mutations may also be negative predictive biomarkers for anti-EGFR therapy. Here we present the description of a new series in which has been performed the analysis of mutations in KRAS and NRAS genes. Methods: Samples of 108 patients with colorectal cancer were used. DNA were extracted from formalin fixed paraffin embedded samples using COBAS DNA sample preparation kit (Roche). Mutation status was determined by targeted pyrosequencing with N/K RAS Pyro Kit (Qiagen). Results: Sixty-four out of 108 samples were wild type (WT) both for KRAS and NRAS. Thirty-five patients presented mutation on KRAS (27 in codon 12, 4 in 13, 1 in 117, and 3 in 146). The rest of the patients (9) presented alterations in NRAS (1 in codon 12, 3 in 13, and 5 in 61). Conclusion: Our results closely resemble those published to date,2 but due to small sample size are not fully comparable. Further studies must be done to determine the real distribution of the new RAS mutations. ReferencesDouillard JY, Oliner KS, Siena S, et al. Panitumumab-FOLFOX4 treatment and RAS mutations in colorectal cancer. N Engl J Med 2013; 369: 1023–34.Oliner KS, Douillard JY, Siena S, et al. Analysis of KRAS/NRAS and BRAF mutations in the phase III PRIME study of panitumumab (pmab) plus FOLFOX versus FOLFOX as first-line treatment (tx) for metastatic colorectal cancer (mCRC). J Clin Oncol 2013; 31 (Suppl): Abstr 3511.