Immune checkpoint inhibitors for POLE or POLD1 proofreading-deficient metastatic colorectal cancer.

BACKGROUND: POLE and POLD1 proofreading deficiency (POLE/D1pd) define a rare subtype of ultramutated metastatic colorectal cancer (mCRC; over 100 mut/Mb). Disease-specific data about the activity and efficacy of immune checkpoint inhibitors (ICIs) in POLE/D1pd mCRC are lacking and it is unknown whether outcomes may be different from mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) mCRCs treated with ICIs. PATIENTS AND METHODS: In this global study, we collected 27 patients with mCRC harboring POLE/D1 mutations leading to proofreading deficiency and treated with anti-programmed cell death-ligand 1 alone +/- anti-cytotoxic T-lymphocyte antigen-4 agents. We collected clinicopathological and genomic characteristics, response, and survival outcomes after ICIs of POLE/D1pd mCRC and compared them with a cohort of 610 dMMR/MSI-H mCRC patients treated with ICIs. Further genomic analyses were carried out in an independent cohort of 7241 CRCs to define POLE and POLD1pd molecular profiles and mutational signatures. RESULTS: POLE/D1pd was associated with younger age, male sex, fewer RAS/BRAF driver mutations, and predominance of right-sided colon cancers. Patients with POLE/D1pd mCRC showed a significantly higher overall response rate (ORR) compared to dMMR/MSI-H mCRC (89% versus 54%; P = 0.01). After a median follow-up of 24.9 months (interquartile range: 11.3-43.0 months), patients with POLE/D1pd showed a significantly superior progression-free survival (PFS) compared to dMMR/MSI-H mCRC [hazard ratio (HR) = 0.24, 95% confidence interval (CI) 0.08-0.74, P = 0.01] and superior overall survival (OS) (HR = 0.38, 95% CI 0.12-1.18, P = 0.09). In multivariable analyses including the type of DNA repair defect, POLE/D1pd was associated with significantly improved PFS (HR = 0.17, 95% CI 0.04-0.69, P = 0.013) and OS (HR = 0.24, 95% CI 0.06-0.98, P = 0.047). Molecular profiling showed that POLE/D1pd tumors have higher tumor mutational burden (TMB). Responses were observed in both subtypes and were associated with the intensity of POLE/D1pd signature. CONCLUSIONS: Patients with POLE/D1pd mCRC showed more favorable outcomes compared to dMMR/MSI-H mCRC to treatment with ICIs in terms of tumor response and survival.

Primary and metastatic brain cancer genomics and emerging biomarkers for immunomodulatory cancer treatment.

Recent studies with immunomodulatory agents targeting both cytotoxic T-lymphocyte protein 4 (CTLA4) and programmed cell death 1 (PD1)/programmed cell death ligand 1 (PDL1) have shown to be very effective in several cancers revealing an unexpected great activity in patients with both primary and metastatic brain tumors. Combining anti-CTLA4 and anti-PD1 agents as upfront systemic therapy has revealed to further increase the clinical benefit observed with single agent, even at cost of higher toxicity. Since the brain is an immunological specialized area it's crucial to establish the specific composition of the brain tumors' microenvironment in order to predict the potential activity of immunomodulatory agents. This review briefly summarizes the basis of the brain immunogenicity, providing the most updated clinical evidences in terms of immune-checkpoint inhibitors efficacy and toxicity in both primary and metastatic brain tumors with the final aim of defining potential biomarkers for immunomodulatory cancer treatment.

2-methoxyestradiol induces morpho-functional changes and impairs the microtubular system in mouse neuroblastoma and rat glioma cells.

2-Methoxyestradiol (2ME), a metabolite deriving from 17-beta estradiol, is a well-established antiangiogenic, apoptotic and antiproliferative agent in cell cultures and animal models. 2ME may also exert its cytotoxic activity by interacting with tubulin and by causing an impairment of the microtubular system. The aim of this study was to investigate the relative effectiveness of 2ME on mouse neuroblastoma (C1300) and rat glioma (C6) cell lines in inducing morpho-functional changes and alteration of the microtubular system physiology. Cells, cultured in a medium supplemented with increasing 2ME micromolar concentrations, were submitted to morphological investigations, MTT assay and western blot analysis. 2ME-exposed cell lines displayed in comparison with control cells, morpho-functional changes such as reduction in cell number, a globular/shrunken shape, retraction or absence of cytoplasmic processes, inhibition of cell growth and cell decreased viability. Interestingly, all changes detected were more evident in C1300 cells than in C6 cells. Western blot analysis showed that the total and the tyrosinated a-tubulin expression was reduced more intensely in the C1300 than in C6 cells; whereas the acetylated a-tubulin expression did not significantly decrease in either cell lines. Results demonstrate that 2ME is more effective in neural cells than in glial cells. The alteration of total and tyrosinated a-tubulin expression suggests that 2ME effectiveness could be strictly related to an impairment of microtubule system physiology resulting in morpho-functional changes, block of mitosis and cell death.

D-glucose induces microtubular changes in C1300 neuroblastoma cell line through the incorporation of 3-nitro-L-tyrosine into tubulin.

The microtubular network of neurons is involved in several functions such as formation and tropism of cellular processes, cell division and intracellular transport. A lot of evidences testify that the microtubular network of neurons can be impaired by oxidative stress. A condition of oxidative stress is often possible when D-glucose overloads its metabolic pathway, resulting in an increase in reactive oxygen species and subsequent neurological disorders. The aim of this work was to check in undifferentiated mouse neuroblastoma cells (C1300) the possible oxidative effects of D-glucose on microtubules. Using a concentration of 110mM D-glucose, cell morphology, growth rate, viability and catalase activity were seriously altered. Noteworthy, an increase in 3-nitro-L-tyrosine and a downregulation of tubulins was found in D-glucose-exposed cells, whereas another cytoskeletal proteins, namely actin, did not show any changes. In conclusion, microtubular network can be impaired by D-glucose through specific nitrosative effects, suggesting a possible mechanism at the basis of hyperglycemia-induced neuronal damage.

Testosterone induces neuroprotection from oxidative stress. Effects on catalase activity and 3-nitro-L-tyrosine incorporation into alpha-tubulin in a mouse neuroblastoma cell line.

3-nitro-L-tyrosine is formed by nitric oxide following different pathways such as NADPH oxidase, xanthine oxidase or glutamate NMDA receptor activation and is involved in the pathology of different neurological disorders. Unlike estradiol, a neuroprotective role of androgens against oxidative cell injury has not been fully investigated. This work targets the possible effects of testosterone on neuroblastoma cells exposed to 3-nitro-L-tyrosine. C1300 mouse undifferentiated neuroblastoma cells exposed to 3-nitro-L-tyrosine were cultured in the presence of testosterone. Morphological examination, proliferation and nuclear viability assays were performed. The expression of tyrosinated alpha-tubulin and incorporation of 3-nitro-L-tyrosine into protein were also estimated. Cells exposed to 3-nitro-L-tyrosine showed globular shape, reduced cytoplasmic processes and growth inhibition in comparison with controls. When testosterone was added to the medium, these changes were not evident. In addition, testosterone induced an upregulation of tyrosinated alpha-tubulin, a marker of neuronal plasticity, and a decrease in 3-nitro-L-tyrosine incorporation into tubulin. Our results suggest that testosterone exposure can diminish 3-nitro-L-tyrosine toxic effects on the morphology and growth rate of neuroblastoma cells. The upregulation of tyrosinated alpha-tubulin in testosterone-exposed cells would be consistent with concurrent plasticity events. Failure in alpha-tubulin nitration detected in cells exposed to both 3-nitro-L-tyrosine and testosterone, may support the idea that testosterone interferes with 3-nitro-L-tyrosine protein incorporation. Moreover, testosterone-induced neuroprotection likely entails a linkage with the androgen receptor as is suggested by the flutamide-induced inhibition of the hormone activity. Finally, the neuroprotective effects of testosterone in neuroblastoma cells could deal with the cellular antioxidant defence system, as shown by testosterone-induced increase in catalase activity.

Helicobacter pylori antibodies in areas of Italy at varying gastric cancer risk.

In a survey of 930 adults aged 35-74 years randomly sampled from the general population of four areas of Italy, two at low and two at high risk for gastric cancer, plasma levels of Helicobacter pylori IgG antibodies were assayed in order to investigate associations with the geographical distribution of gastric cancer and other dietary and life-style factors, as assessed by personal interview. H. pylori positivity (antibody titer above or equal to 10 micrograms/ml), 45% overall, increased with age and was inversely associated with social class but showed little geographical variation or association with dietary variables and blood nutrients. H. pylori positivity was also associated with increased blood levels of pepsinogens, particularly pepsinogen II. The authors discuss these findings in relation to those from a previous case-control study of gastric cancer in the same areas.

Plasma pepsinogens, nutrients, and diet in areas of Italy at varying gastric cancer risk.

In a survey of 930 adults aged 35-74 years randomly sampled from the general population of four areas of Italy at different risks for gastric cancer (GC), plasma levels of pepsinogens (PGI and PGII) and fat-soluble vitamins were assayed. Pepsinogen levels were used to identify individuals with chronic atrophic gastritis (CAG). Severe CAG (PGI < or = 20 pg/liter) affected 5.8% of the population, but the prevalence rose with increasing age and declining social class. Severe CAG was 5 times more common in areas with high compared to low rates of GC. Risk also rose with increasing consumption of salted/dried fish but was inversely related to dietary intake of beta-carotene and to plasma retinol and cholesterol levels. The prevalence of moderate CAG (PGI > 20 pg/liter, but PGI/PGII < or = 2.9) was 6.3%. Moderate CAG was also related to age and social class and increased 1.8-fold in areas where GC rates were high, but was not strongly associated with diet or plasma nutrients. The authors discuss these findings in relation to those from a previous case-control study of GC in the same areas.

Variations of plasma CEA levels in gynecologic malignancies: incidence, significance and changes occurring in response to therapy.

Serial carcinoembryonic antigen (CEA) radioimmunoassay were performed on 84 patients with primary gynecologic malignancies using the double antibody method. A positive correlation between the marker levels and tumor stage was found in the subjects with invasive disease: the incidence of positive values is appreciably higher in the advanced stages of malignancies. The surgical resection normalized the pretreatment elevated levels: fluctuating CEA values were observed during chemio or radiotherapy. The findings of this investigation support the theory of the limited usefulness of the CEA levels as a diagnostic test in patients with gynecologic cancer. The determination would appear to be helpful only in case with an elevated plasma values at the time of diagnosis. The CEA test seems to indicate that serial plasma assays can be of large interest in the follow-up of this women and for the second-look surgery, chemio and radiant therapy.

[Notes on the development of treatment protocols in epithelial head and neck tumors]. / Nota sintetica sulla evoluzione dei protocolli terapeutici nei tumori epiteliali testa-collo.

Observation of data relating to a lengthy period of clinical activity in the field of head and neck tumours has provided the opportunity for drawing a number of conclusions as regards the prophylaxis of such situations as well as chemo-radiosurgical operating schedules. As regards prophylaxis, particular stress is laid on the worrying number of late diagnoses. In matters of therapy, emphasis is laid on the need for more specific immunological assessments in patients, to guide the physician's decision. The interest of extensive chemotherapeutic experience, which is especially abundant in non-surgical patients, is stressed for the purpose of getting ready for all future developments in the field.

Antispasmodic activity of tert-aminoalkylderivatives of 3-benzyl-, 3-benzylaza- and 3-benzyldiazaquinoxalinones.

Tert-aminoalkylderivatives of quinoxalin-2-ones, aza- and diazaquinoxalin-2-ones bearing in position 3 a benzyl group were assayed to evaluate the antispasmodic activity. The tested compounds exhibited moderate aspecific antispastic properties that do not warrant further investigation.

Synthesis and choleretic activity of 3-[2-(3-R', 4-R'', 5-R'''-benzyl)-5-R-benzimidazol-1-yl]-butanoic acids.

On the ground of the evidentiated choleretic activity of 3-[2-benzylbenzimidazol-1-yl]butanoic acid, 28 new acids were prepared in order to evaluate the influence of suitable substitutions in either C5 of heteroring or C3', C4', C5' of benzyl group in position 2 on the choleretic activity. Pharmacological results after i.v. administration of 0.5 mmol/Kg in rats confirmed a general high choleretic activity that in eleven cases showed during the first 4 hours an increase of bile volume higher than 80%, that is superior to that produced by dehydrocholic acid. Only in a few cases the bile volume increase was less than 37% of basal value.

Effect of thyrotropin-releasing hormone in pituitary-thyroid system function infertile women after surgical castration.

The Authors evaluate the effect of surgical castration on thyroid function of fertile women, and the response of TSH to TRH before and 30 days after castration.

[Silica, silicosis, and lung cancer: analysis if the literature and mortality studies of minor workers in Sardinia]. / Silice, silicosi e cancro del polmone: analisi della letteratura e studi di mortalità in minatori della Sardegna.

Starting from a short review of the recent epidemiological studies available in the international literature concerning the association between silica, silicosis and lung cancer, the results of two mortality studies performed in Sardinia are reported. The first study concerns a 20-year follow-up of 1741 miners employed in 1973 in two metalliferous Sardinian mines. In the second study the cause specific mortality of 724 patients with silicosis, firstly diagnosed by standard chest x-ray between 1964 and 1970 in our Institute, has been analysed by a cohort study extended to December 31, 1997. The findings indicate that the slight increased lung cancer mortality observed in these cohorts, more than to the severity of radiological silicosis or to the entity of the cumulative exposure to crystalline silica dust in itself, was significantly associated to other risk factors as cigarette smoking, airflow obstruction and radon-daughters exposure in underground mines.

Distribution of SNAP25, VAMP1 and VAMP2 in mature and developing deep cerebellar nuclei after estrogen administration.

Synaptosomal-associated protein of 25kDa (SNAP25), vesicle-associated membrane protein 1 (VAMP1) and 2 (VAMP2) are components of soluble N-ethylmaleimide-sensitive fusion attachment protein receptors (SNARE) complex which is involved in synaptic vesicle exocytosis, a fundamental step in neurotransmitter release. SNARE expression in cerebellum correlates with specific neurotransmitter pathways underlying synaptic diversification and defined synaptic properties. In this study we firstly characterized the distribution of SNAP25, VAMP1 and VAMP2 in the nerve terminals of a defined cerebellar region, the deep cerebellar nuclei (DCN), of adult and newborn rats. Then, given the pivotal role of estradiol (E2) in the synaptic organization of the cerebellar circuitry in early postnatal life, we examined whether administration of E2 in the newborn DCN affected synaptic density and changed the distribution of the presynaptic proteins SNAP25, VAMP1 and VAMP2, together with post synaptic density protein 95 (PSD95). Results showed that: (1) distribution of SNAP25, VAMP1 and VAMP2 in adult DCN differs significantly from that found in newborn DCN; (2) administration of E2 in the newborn DCN affected synaptic density and also changed the distribution of the pre- and postsynaptic proteins. The differential distribution of SNAP25, VAMP1 and VAMP2 in nerve terminals of adult and newborn rats may correlate with specific stages of neuronal phenotypic differentiation. The effects of E2 on SNAP25, VAMP1, VAMP2, PDS95 and synaptic density suggest that pre- and postsynaptic proteins are under estrogenic control during development and that synaptic maturation can also be related with the activity of this steroid.

Immunohistochemical localisation and molecular expression of the steroidogenic enzyme cytochrome P450 17α-hydroxylase /C(17,20)-lyase in the vestibular nuclei of adult male rats.

Many biologically active neurosteroids, including dehydroepiandrosterone (DHEA), are synthesised in the brain. DHEA is a potent endogenous modulator of several neuronal functions, and alterations of DHEA are correlated with various neurobiological deficits. The cytochrome P450 17α-hydroxylase/C(17,20)-lyase (P450C(17) ) plays a pivotal role in the synthesis of DHEA from pregnenolone and progesterone. We investigated the immunohistochemical localisation and molecular expression of P450C(17) in the superior, lateral, medial and inferior vestibular nuclei (VCN) of adult male rats by western blotting and indirect immunofluorescence analysis. Immunoreactive P450C(17) was widely distributed in all VCN and the expression of P450C(17) was confirmed by western blot analysis. The present study demonstrates, for the first time, the presence and anatomical distribution of P450C(17) in the VCN. Given that neurosteroids can modulate neuronal activities in the medial vestibular nucleus, DHEA synthesised in the VCN may play an important role in the control of specific activities at this level.