PD-1 and PD-L1 expression in molecularly selected non-small-cell lung cancer patients.

BACKGROUND: Agents targeting programmed death-1 receptor (PD-1) and its ligand (PD-L1) are showing promising results in non-small-cell lung cancer (NSCLC). It is unknown whether PD-1/PD-L1 are differently expressed in oncogene-addicted NSCLC. METHODS: We analysed a cohort of 125 NSCLC patients, including 56 EGFR mutated, 29 KRAS mutated, 10 ALK translocated and 30 EGFR/KRAS/ALK wild type. PD-L1 and PD-1 expression were assessed by immunohistochemistry. All cases with moderate or strong staining (2+/3+) in >5% of tumour cells were considered as positive. RESULTS: PD-1 positive (+) was significantly associated with current smoking status (P=0.02) and with the presence of KRAS mutations (P=0.006), whereas PD-L1+ was significantly associated to adenocarcinoma histology (P=0.005) and with presence of EGFR mutations (P=0.001). In patients treated with EGFR tyrosine kinase inhibitors (N=95), sensitivity to gefitinib or erlotinib was higher in PD-L1+ vs PD-L1 negative in terms of the response rate (RR: P=0.01) time to progression (TTP: P<0.0001) and survival (OS: P=0.09), with no difference in PD1+ vs PD-1 negative. In the subset of 54 EGFR mutated patients, TTP was significantly longer in PD-L1+ than in PD-L1 negative (P=0.01). CONCLUSIONS: PD-1 and PD-L1 are differentially expressed in oncogene-addicted NSCLC supporting further investigation of specific checkpoint inhibitors in combination with targeted therapies.

Re-differentiation of thyroid carcinoma cell lines treated with 5-Aza-2'-deoxycytidine and retinoic acid.

We studied cell growth rate, mechanisms of growth inhibition, phenotype re-differentiation, expression of RARalpha, beta, gamma and differentiation thyroid genes before and after combined treatment with 5-Aza-CdR and RA (5-Aza/RA) of human thyroid carcinoma cell lines (FRO, WRO, TT). Furthermore, the activity and localization of the re-expressed sodium-iodide-symporter (NIS) protein was analyzed. After 5-Aza/RA treatment, all cell lines showed a significant reduction in cell growth. This was associated with apoptosis in the TT, with inhibition of cell proliferation in the WRO, and with cell cycle impairment in FRO and WRO. FRO and WRO treated with 5-Aza/RA lost the ability to grow in soft agar. FRO re-expressed thyroid transcription factor-1 and thyroglobulin, TT and WRO re-expressed PAX-8 and FRO and TT re-expressed RARbeta and NIS mRNA. Despite this expression, they were unable to take up iodine: a cytoplasmic localization of NIS protein was demonstrated in FRO. In conclusion, besides a significant reduction in cell growth rate and in the ability to grow in soft agar, treatment with 5-Aza/RA partially re-differentiated FRO and induced expression of NIS mRNA and protein in FRO and TT, but this treatment was unable to restore the functional activity of NIS, likely because it was located into the cytoplasm without reaching the plasma membrane.

Retinoic acid receptor beta2 re-expression and growth inhibition in thyroid carcinoma cell lines after 5-aza-2'-deoxycytidine treatment.

The treatment of both undifferentiated and de-differentiated thyroid tumors, which are unresponsive to radioiodine, represents one of the biggest challenges for thyroidologists. The aim of the present study was to investigate in vitro the methylation status of retinoic acid receptors (RAR)beta2 promoter and the effect of the demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on 5 human thyroid cancer cell lines. The methylation status of RARbeta2 promoter was analyzed by methylation-specific PCR. The effect of 5-Aza-CdR on cell growth and apoptosis was evaluated by cell counting, enzymelinked immunosorbent assay tests and fluorescence-activated cell sorting analysis, while the effect on the expression of RAR and thyroid-specific genes was measured by qualitative and quantitative RT-PCR. Methylation of RARbeta2 promoter was present only in ARO cells. 5-Aza-CdR determined growth inhibition in all cell lines, probably due to apoptosis in WRO, NPA, and ARO cells, and to inhibition of DNA synthesis in TT cells. Treatment with 5-Aza-CdR induced the expression of RARbeta mRNA in ARO and FRO cells, a slight increase of the expression of Tg, TPO and thyroid trancription factor 1 (TTF-1) mRNA and the new expression of low levels of NIS in TT cells. A significant increase of TTF-1 mRNA in FRO cells was also observed. In this study we demonstrated that RARbeta2 promoter was methylated in ARO cell line. However, the 5-Aza-CdR treatment induced RARbetamRNA expression not only in ARO but also in FRO and TT cell lines, whose RARbeta2 promoter was unmethylated. A significant reduction of cell growth, but not cell re-differentiation, was also observed after 5-Aza-CdR treatment.

PS-341 (Bortezomib) inhibits proliferation and induces apoptosis of megakaryoblastic MO7-e cells.

PS-341 (Bortezomib) is a dipeptide boronic acid proteasome inhibitor with antitumor activity that induces apoptosis in different human cancer cell lines. We investigated effects of PS-341 (Bortezomib) on cell proliferation, cell cycle progression, induction of apoptosis and differentiation in a megakaryoblastic (MO7-e) cell line. PS-341 was able to retain NF-kappaB in the cytoplasm and inhibit cell growth (IC(50)=22.5 nM), in a dose/time-dependent way. This anti-proliferative activity resulted to be lineage-specific, because other leukemic cell lines (KG1a, K562/R7, HL60/DNR) were unaffected by the PS-341 treatment. Moreover, PS-341 in MO7-e induced a significant pro-apoptotic effect from 10 nM concentration (40% versus 12% in the control, p<0.05). On the other hand, at lower concentration (5 nM), Bortezomib blocked cell cycle in the G2 phase. Finally, this compound was able to down-regulate WT1 expression. No significant effects on cell differentiation were found. Because a spontaneous NF-kappaB activation has been reported in megakaryocytes from patients affected by myeloproliferative disorders, Bortezomib would so be an attractive therapeutic tool for these malignancies, including essential thrombocythemia or idiopathic myelofibrosis. Preliminary data show an inhibiting activity of Bortezomib in the megakaryocytic colonies formation. Finally, also down-regulation of the WT1 gene Bortezomib-driven could be relevant, because of the role that this gene would play in the pathogenesis of acute and chronic myeloproliferative disorders.

Dimensions of social adjustment following psychiatric hospitalization: a six-month follow-up study.

This study describes a prospective six-month follow-up study of previously hospitalized psychiatric patients. Data are presented on social functioning and symptomatology derived from self-report scales administered to 60 patients. there was significant improvement in both symptomatology and social functioning on follow-up for all groups with the exception of the schizophrenic group, who exhibited little or no change. Residual change scores revealed more than expected change in 71 percent of schizophreniform patients, 70 percent of nondepressed patients, 53 percent of depressed patients, but only 20 percent of schizophrenic patients. These findings suggest that most patients sustain improvement after discharge and that many schizophreniform patients may have good prognoses.

The outpatient treatment of heroin addicts with methadone: a two- to five-year follow-up study.

This study is a two- to five-year follow-up of 40 patients who were discharged from a methadone maintenance program. Both "graduates" and prematurely discharged patients were surveyed. Forty-five percent of patients had been in treatment for one year or more. Outcome evaluation was based on a reduction of illicit drug use, an increase in social stability, and a decline in criminal activity. Eighteen of 40 clients were located and interviewed. Seven (39 percent) were found to be either drug free or using only prescribed methadone, and five had substantially reduced their drug use. Relapse to illicit drug use was associated with a pre-treatment history of poly-drug use, poorer work and arrest records, and a greater number of previous treatments. Abstinence appeared to be a realistic goal for only a few. The importance of retaining patients in treatment longer is discussed.

The dexamethasone suppression test in suicidal patients with unipolar depression.

Of 49 newly hospitalized patients who met Research Diagnostic Criteria for primary unipolar depression and were given the dexamethasone suppression test (DST), significantly more patients admitted for suicide attempts than nonsuicidal patients had abnormal DSTs. All five patients who made subsequent suicide attempts (one completed) within 6 months after admission had had abnormal DSTs. These findings suggest that the tendency of endogenously depressed patients to attempt suicide is exacerbated by an underlying neurobiological disorder reflected by limbic-hypothalamic dysregulation and that depressed suicidal patients with abnormal DSTs represent a high-risk group for recurrence of suicidal behavior.

The dexamethasone suppression test in adolescent psychiatric inpatients.

The dexamethasone suppression test (DST) was administered to 120 adolescent psychiatric patients at the time of hospitalization, and cortisol levels were measured at 4:00 p.m. and 11:30 p.m. on the day after a 1-mg oral dose of dexamethasone was given. Failure to suppress serum cortisol (i.e., cortisol level less than 5 micrograms/dl) was noted in 25 patients: 7 of 17 patients who met DSM-III criteria for major depressive disorder, 7 of 38 patients with dysthymic disorder, 7 of 47 patients with conduct disorder, and 4 of 15 schizophreniform patients. The predictive value of the DST for major depressive disorder was only 28%. Although adolescent patients with abnormal DSTs may eventually develop affective symptoms consistent with a major depressive disorder, the DST did not discriminate between major depression and other psychiatric diagnoses in these hospitalized adolescents.

Assessment of suicide risk: some limitations in the prediction of infrequent events.

Efforts to predict suicide and attempted suicide generally have yielded equivocal results. Neither single signs, standard psychological tests, specially devised tests, clinical judgments nor scales are found to be able to predict suicide at useful levels. Prediction of suicide is hampered by the relative rarity of this behavior together with the inadequate specificity of clinical characteristics and antecedent events. No highly unique descriptive attributes of suicide committers have been discovered. Within these existing limitations, a set of potential predictor variables are presented that may result in improved prediction of suicidal risk.

An intervention to reduce the rate of hospital discharges against medical advice.

The initiation of a patient advocacy program in a private psychiatric hospital in 1980 was effective in significantly reducing the rate of hospital discharges against medical advice relative to the rate in 1979 and in 1978. There was also a concomitant increase in the rate of clinically approved discharges relative to that in 1979 and in 1978. The authors view the patient advocate as an objective intermediary who represents an acknowledgment of patients' rights and affords patients an opportunity to have some autonomy and an impact on the hospital system.