Determinants of activity and efficacy of anti-PD1/PD-L1 therapy in patients with advanced solid tumors recruited in a clinical trials unit: a longitudinal prospective biomarker-based study.

Immune-checkpoint inhibitors (ICI) have revolutionized the therapeutic landscape of cancer. However, optimal patient selection is still an unmet need. One-hundred-forty-six patients with metastatic cancer candidates to ICI at the Hospital Clinic of Barcelona Clinical Trials Unit were prospectively recruited in this observational study. Blood samples were collected at different timepoints, baseline LIPI score calculated and pre-ICI archived tissues retrieved to evaluate PD-L1, tumor-infiltrating lymphocytes (TILs) and PD1 mRNA levels. Tumor assessments were centrally reviewed by RECIST 1.1 criteria. Associations with overall response rates (ORR), durable clinical benefit (DCB), progression-free survival (PFS) and overall survival (OS) were performed with univariable/multivariable logistic and Cox regressions, where appropriate. At a median follow-up of 26.9 months, median PFS and OS were 2.7 and 12.9 months. Response rates were 17.8% with duration of response (DOR) of 4.4 months. LIPI score was independently associated with PFS (p = 0.025) and OS (p < 0.001). Immunotherapy-naïve status was independently associated with better PFS (p = 0.005). Time-to-best response (TTBR) and ORR (p < 0.001 both) were associated with better OS at univariate analysis. PFS and DOR were moderately correlated with OS (p < 0.001 both). A PD-L1 10% cut-off detected worse/best responders in terms of ORR (univariate p = 0.011, multivariate p = 0.028) and DCB (univariate p = 0.043). PD1 mRNA levels were strikingly associated to complete responses (p = 0.021). To resume, in our prospective observational pan-cancer study, baseline LIPI score, immunotherapy-naïve status, cancer type and RT before starting ICI were the most relevant clinical factors independently correlated with immunotherapy outcomes. Longer TTBR seemed to associate with better survival, while PD1 mRNA and PD-L1 protein levels might be tumor-agnostic predictive factors of response to ICI and should be furtherly explored.

Activity of silver nanoparticles on prokaryotic cells and Bothrops jararacussu snake venom.

Nanoparticle-conjugated venom-toxins of venomous animals and its therapeutic efficacy against emerging or neglecting diseases is a promising strategy. In this study, silver nanoparticles (AgNPs ∼50 nm, 0.081 mg mL-1) were studied against the neuromuscular blockade, myotoxic effects induced by Bothrops jararacussu venom (60 µg mL-1) and also against prokaryotic cells. The neurotoxicity was evaluated on ex vivo mouse phrenic nerve-diaphragm using traditional myographic technique, able to obtain functional contractile responses and to check the neurotransmission. The myotoxicity on mammalian cells was evaluated in muscles resulting from pharmacological assays using routine histological techniques and light microscopy. The toxicity to prokaryotic cells was evaluated on Salmonella typhimurium TA100 without metabolic activation. The in vitro preincubation model between AgNPs and venom was enough to abolish toxic effects of B. jararacussu venom, but mammalian cells were highly sensitive to AgNPs more than prokaryotic cells, by acting as dose-independently and dose-dependently parameters, respectively. These results allowed us to conclude that AgNPs showed promising activity as antivenom agent but for its safer use, the toxicity should be evaluated on experimental animals.

Are Silver Nanoparticles Useful for Treating Second-Degree Burns? An Experimental Study in Rats.

Purpose: In this work, the potential usefulness of silver nanoparticles (AgNPs) for treating burn wounds was examined. Methods: Second-degree burns were induced in male Wistar rats by touching the skin with a heated (70°C) metallic device for 10 s, after which the animals were randomly allocated to one of two groups: control (n=8, treated with sterile saline) and experimental (n=8, treated with AgNPs, 0.081 mg/mL; 50 µL applied to the burn surface). Seven, 14, 21 and 28 days after lesion induction two rats from each group were killed and blood samples were collected for a complete blood count and to assess oxidative stress. The livers were examined macroscopically and skin samples were collected for histological analysis. Results: Macroscopically, wound healing and skin remodeling in the experimental group were similar to the saline-treated rats. Likewise, there were no significant differences in the histological parameters between the two groups. However, treatment with AgNPs caused a persistent reduction in white blood cell (WBC) counts throughout the experiment, whereas platelet counts increased on days 7 and 28 but decreased on days 14 and 21; there was also an increase in the blood concentration of reduced glutathione on day 7 followed by a decrease on days 21 and 28. There were no significant changes in blood glutathione peroxidase (GSH-Px) and catalase (CAT) activities or in the serum concentration of thiobarbituric acid reactive substances. Conclusion: The findings of this study raise questions about the potential transitory effects of AgNPs based on the changes in WBC and platelet counts, blood glutathione concentrations and macroscopic hepatic alterations.

Evaluation of Betulin Mutagenicity by Salmonella/Microsome Test.

Purpose: Betulin is a pentacyclic triterpene found in the outer barks of innumerous plants. This secondary metabolite is easily isolated from plants with the major interest in converting it to betulinic acid, which pharmacological properties were much more exploited than betulin. But, investments in the own betulin have been grown since no chemical step is necessary. In this study we focused the precursor betulin in order to evaluate its mutagenicity by Salmonella/microsome assay (Ames test). Methods: The Ames test was carried out using a commercial betulin exposed to Salmonella typhimurium strains TA98, TA100, TA102, and TA97a, in experiments with (+S9) and without (-S9) metabolic activation. Results: Betulin was unable to increase the number of revertants (+S9 and -S9 metabolic activation) showing the absence of any mutagenic effect by Ames test. Conclusion: This study allowed attribute safety to betulin being important for exploiting its pharmacological uses.

A personalized preclinical model to evaluate the metastatic potential of patient-derived colon cancer initiating cells.

PURPOSE: Within the aim of advancing precision oncology, we have generated a collection of patient-derived xenografts (PDX) characterized at the molecular level, and a preclinical model of colon cancer metastasis to evaluate drug-response and tumor progression. EXPERIMENTAL DESIGN: We derived cells from 32 primary colorectal carcinomas and eight liver metastases and generated PDX annotated for their clinical data, gene expression, mutational, and histopathological traits. Six models were injected orthotopically into the cecum wall of NOD-SCID mice in order to evaluate metastasis. Three of them were treated with chemotherapy (oxaliplatin) and three with API2 to target AKT activity. Tumor growth and metastasis progression were analyzed by positron emission tomography (PET). RESULTS: Patient-derived cells generated tumor xenografts that recapitulated the same histopathological and genetic features as the original patients' carcinomas. We show an 87.5% tumor take rate that is one of the highest described for implanted cells derived from colorectal cancer patients. Cecal injection generated primary carcinomas and distant metastases. Oxaliplatin treatment prevented metastasis and API2 reduced tumor growth as evaluated by PET. CONCLUSIONS: Our improved protocol for cancer cell engraftment has allowed us to build a rapidly expanding collection of colorectal PDX, annotated for their clinical data, gene expression, mutational, and histopathological statuses. We have also established a mouse model for metastatic colon cancer with patient-derived cells in order to monitor tumor growth, metastasis evolution, and response to treatment by PET. Our PDX models could become the best preclinical approach through which to validate new biomarkers or investigate the metastatic potential and drug-response of individual patients.