PD-L1 overexpression induces STAT signaling and promotes the secretion of pro-angiogenic cytokines in non-small cell lung cancer (NSCLC).

BACKGROUND: Monoclonal antibodies (ICI) targeting the immune checkpoint PD-1/PD-L1 alone or in combination with chemotherapy have demonstrated relevant benefits and established new standards of care in first-line treatment for advanced non-oncogene addicted non-small cell lung cancer (NSCLC). However, a relevant percentage of NSCLC patients, even with high PD-L1 expression, did not respond to ICI, highlighting the presence of intracellular resistance mechanisms that could be dependent on high PD-L1 levels. The intracellular signaling induced by PD-L1 in tumor cells and their correlation with angiogenic signaling pathways are not yet fully elucidated. METHODS: The intrinsic role of PD-L1 was initially checked in two PD-L1 overexpressing NSCLC cells by transcriptome profile and kinase array. The correlation of PD-L1 with VEGF, PECAM-1, and angiogenesis was evaluated in a cohort of advanced NSCLC patients. The secreted cytokines involved in tumor angiogenesis were assessed by Luminex assay and their effect on Huvec migration by a non-contact co-culture system. RESULTS: PD-L1 overexpressing cells modulated pathways involved in tumor inflammation and JAK-STAT signaling. In NSCLC patients, PD-L1 expression was correlated with high tumor intra-vasculature. When challenged with PBMC, PD-L1 overexpressing cells produced higher levels of pro-angiogenic factors compared to parental cells, as a consequence of STAT signaling activation. This increased production of cytokines involved in tumor angiogenesis largely stimulated Huvec migration. Finally, the addition of the anti-antiangiogenic agent nintedanib significantly reduced the spread of Huvec cells when exposed to high levels of pro-angiogenic factors. CONCLUSIONS: In this study, we reported that high PD-L1 modulates STAT signaling in the presence of PBMC and induces pro-angiogenic factor secretion. This could enforce the role of PD-L1 as a crucial regulator of the tumor microenvironment stimulating tumor progression, both as an inhibitor of T-cell activity and as a promoter of tumor angiogenesis.

Hormonal and metabolic effects of neuroglucopenia.

We examined the role of central neuroglucopenia, induced by intracerebroventricular (i.c.v.) administration of 2-deoxyglucose (2-DG), on glucose and amino acid kinetics in conscious dogs. Group 1 received i.c.v. 2-DG at 2.5 mg.kg-1 x min-1 for 15 min. Group 2 received an equal intravenous (i.v.) amount of 2-DG. In the i.c.v. group, plasma glucose levels rose from 106 +/- 4 mg/dl to a peak of 204 +/- 12 mg/dl by 90 min. Blood lactate increased from 689 +/- 1 to 2,812 +/- 5 mumol/l and blood alanine not change from basal (256 +/- 41 mumol/l). The rate of hepatic glucose production, determined isotopically, was increased 2-fold over basal (P < 0.01). Significant increases (P < 0.001) over basal were also noted in plasma epinephrine, norepinephrine, insulin, glucagon and cortisol. Leucine rate of appearance (Ra) showed a 30% decrease from basal to 2.4 +/- 0.05 mumol.kg-1 x min-1 (P < 0.01). In group 2 plasma glucose levels were not altered but plasma cortisol and glucagon showed a modest transient increase above basal (P < 0.05). No significant changes were noted in amino acid kinetics. These findings suggest that periventricular neuroglucopenia, in the absence of peripheral glucose deprivation, is accompanied by hyperglycemia secondary to enhanced hepatic glucose production with decreased glucose utilization and by increased hepatic uptake of gluconeogenic precursors. These, however, were not accompanied by increased whole body proteolysis as was previously seen with generalized glucopenia resulting from insulin-induced hypoglycemia.

The role of the central nervous system in modulating glucose and protein metabolism during insulin-induced hypoglycemia.

We previously established that insulin-induced hypoglycemia is associated with enhanced proteolysis with the GI tract being the major contributor to this response. In the present study we examined whether brain hypoglycemia modulates the changes in amino acid and glucose kinetics during insulin-induced hypoglycemia. Studies were performed in 24 h faster conscious dogs chronically fitted with catheters in the femoral artery, portal vein, hepatic vein, femoral vein and in both carotid and vertebral arteries, and that were also fitted with permanent tracheostomies. Following a 30 min basal period insulin was infused i.v. at 300 mU/kg.h to achieve systemic hypoglycemia averaging 42 +/- 2 mg/dl. In group I (n = 6) both central and systemic (global) hypoglycemia was allowed to develop. The rate of net hepatic glucose output (NHGO) increased by 50% above basal. Plasma leucine increased from 120 +/- 12 mumol/l basally to 154 +/- 30 mumol/l during the last hour of hypoglycemia. The rate of leucine appearance into the plasma compartment (R(a)) increased from 154 +/- 30 to 200 +/- 36 mumol/kg.h and its rate of oxidation increased from 22 +/- 5 to 51 +/- 8 mumol/kg.h, while its non-oxidative rate of disposal (141 +/- 12 mumol/kg.h) did not change. Net leucine balance across the gut was neutral (6 +/- 5 mumol/kg.h) and switched to net output of 96 +/- 24 mumol/kg.h. In group II (n = 5) 10% dextrose in water was infused into both carotid and vertebral arteries to prevent profound CNS glucopenia. This was associated with a drop in NHGO.(ABSTRACT TRUNCATED AT 250 WORDS)