Nanostructured electrocatalysts for organic synthetic transformations.

Organic chemists have made and are still making enormous efforts toward the development of novel green catalytic synthesis. The necessity arises from the imperative of safeguarding human health and the environment, while ensuring efficient and sustainable chemical production. Within this context, electrocatalysis provides a framework for the design of new organic reactions under mild conditions. Undoubtedly, nanostructured materials are under the spotlight as the most popular and in most cases efficient platforms for advanced organic electrosynthesis. This Minireview focuses on the recent developments in the use of nanostructured electrocatalysts, highlighting the correlation between their chemical structures and resulting catalytic abilities, and pointing to future perspectives for their application in cutting-edge areas.

An integrated approach for the benthic habitat mapping based on innovative surveying technologies and ecosystem functioning measurements.

Among marine ecosystems globally, those in the Mediterranean Sea, are facing many threats. New technologies are crucial for enhancing our understanding of marine habitats and ecosystems, which can be complex and resource-intensive to analyse using traditional techniques. We tested, for the first time, an integrated multi-platform approach for mapping the coastal benthic habitat in the Civitavecchia (northern Latium, Italy) coastal area. This approach includes the use of an Unmanned Surface Vehicle (USV), a Remote Operated Vehicle (ROV), and in situ measurements of ecosystem functionality. The echosounder data allowed us to reconstruct the distribution of bottom types, as well as the canopy height and coverage of the seagrass Posidonia oceanica. Our study further involved assessing the respiration (Rd) and net primary production (NCP) rates of P. oceanica and its associated community through in situ benthic chamber incubation. By combining these findings with the results of USV surveys, we were able to develop a preliminary spatial distribution model for P. oceanica primary production (PP-SDM). The P. oceanica PP-SDM was applied between the depths of 8 and 10 m in the studied area and the obtained results showed similarities with other sites in the Mediterranean Sea. Though in the early stages, our results highlight the significance of multi-platform observation data for a thorough exploration of marine ecosystems, emphasizing their utility in forecasting biogeochemical processes in the marine environment.

Conserving Marine Forests: Assessing the Effectiveness of a Marine Protected Area for Cystoseira sensu lato Populations in the Central Mediterranean Sea.

Marine Protected Areas (MPAs) are vital for biodiversity conservation, yet their effectiveness in preserving foundation seaweeds remains understudied. This study investigates the diversity and distribution of Cystoseira sensu lato (including Cystoseira, Ericaria, and Gongolaria, hereafter referred to as Cystoseira s.l.) populations in an MPA located in the central Mediterranean Sea, comparing them with those in two unprotected sites. We hypothesized MPA Cystoseira s.l. populations would display higher diversity and structure compared to outside unprotected sites. Results revealed a total of 19 Cystoseira s.l. species at depths of 0-20 m, with the MPA exhibiting a higher diversity than unprotected sites. Thus, MPAs can play a crucial role in fostering the diversity of Cystoseira s.l. populations. However, no significant differences were observed among the MPA's protection zones, raising questions about the zoning effectiveness. Additionally, our survey uncovered a substantial presence of non-indigenous seaweeds within the MPA. In conclusions, while MPAs improved Cystoseira s.l. diversity compared to unprotected sites, the varying efficacy of protection within MPA zones suggested a necessity for site-specific conservation strategies. The presence of non-indigenous seaweeds emphasizes ongoing challenges. This study provides a baseline for understanding Cystoseira s.l. population dynamics, crucial for future monitoring and conservation efforts in the face of global change.

Biodiversity of Fungi in Freshwater Ecosystems of Italy.

Fungal biodiversity is still mostly unknown and their presence in particular ecosystems such as freshwater habitats is often underestimated. The ecological role that these fungi play in freshwater environments mainly concerns their activity as decomposers of litter and plant material. At present, it is estimated that 3870 species belong to the ecological group of freshwater fungi (13 phyla and 45 classes). In this survey, we provide an overview of the Italian freshwater fungal diversity on the basis of the field and literature data. In the literature, data on freshwater fungi are fragmentary and not updated, focusing mainly on northern Italy where the most important lakes and rivers are present, while data from central and southern Italy (including Sicily and Sardinia) are almost completely ineffective. In particular, Ascomycota are reported in only 14 publications, most of which concern the freshwater environments of Lombardia, Piemonte, and Veneto. Only one publication explores the biodiversity of freshwater Basidiomycota in the wetlands of the Cansiglio forest (Veneto). The field observation allowed for us to identify 38 species of Basidiomycota growing in riparian forest of Italy. However, the number of fungi in freshwater habitats of Italy is strongly underestimated and many species are still completely unknown.

A pan-cancer clinical platform to predict immunotherapy outcomes and prioritize immuno-oncology combinations in early-phase trials.

BACKGROUND: Immunotherapy is effective, but current biomarkers for patient selection have proven modest sensitivity. Here, we developed VIGex, an optimized gene signature based on the expression level of 12 genes involved in immune response with RNA sequencing. METHODS: We implemented VIGex using the nCounter platform (Nanostring) on a large clinical cohort encompassing 909 tumor samples across 45 tumor types. VIGex was developed as a continuous variable, with cutoffs selected to detect three main categories (hot, intermediate-cold and cold) based on the different inflammatory status of the tumor microenvironment. FINDINGS: Hot tumors had the highest VIGex scores and exhibited an increased abundance of tumor-infiltrating lymphocytes as compared with the intermediate-cold and cold. VIGex scores varied depending on tumor origin and anatomic site of metastases, with liver metastases showing an immunosuppressive tumor microenvironment. The predictive power of VIGex-Hot was observed in a cohort of 98 refractory solid tumor from patients treated in early-phase immunotherapy trials and its clinical performance was confirmed through an extensive metanalysis across 13 clinically annotated gene expression datasets from 877 patients treated with immunotherapy agents. Last, we generated a pan-cancer biomarker platform that integrates VIGex categories with the expression levels of immunotherapy targets under development in early-phase clinical trials. CONCLUSIONS: Our results support the clinical utility of VIGex as a tool to aid clinicians for patient selection and personalized immunotherapy interventions. FUNDING: BBVA Foundation; 202-2021 Division of Medical Oncology and Hematology Fellowship award; Princess Margaret Cancer Center.

Warming and nutrient enrichment can trigger seaweed loss by dysregulation of the microbiome structure and predicted function.

Warming and nutrient enrichment are key pervasive drivers of ecological shifts in both aquatic and terrestrial ecosystems, impairing the physiology and survival of a wide range of foundation species. But the underlying mechanisms often remain unclear, and experiments have overlooked the potential effects mediated by changes in the microbial communities. We experimentally tested in the field orthogonal stress combinations from simulated air warming and nutrient enrichment on the intertidal foundation seaweed Cystoseira compressa, and its associated bacterial communities. A total of 523 Amplicon Sequence Variance (ASVs) formed the bacterial community on C. compressa, with 222 ASVs assigned to 69 taxa at the genus level. Most bacteria taxa experienced changes in abundance as a result of additive (65 %) and antagonistic (30 %) interactions between the two stressors, with synergies (5 %) occurring less frequently. The analysis of the predicted bacterial functional profile identified 160 metabolic pathways, and showed that these were mostly affected by additive interactions (74 %) between air warming and nutrient enrichment, while antagonisms (20 %) and synergisms (6 %) were less frequent. Overall, the two stressors combined increased functions associated with seaweed disease or degradation of major cell-wall polymers and other algicidal processes, and decreased functions associated with Quorum Quenching and photosynthetic response. We conclude that warming and nutrient enrichment can dysregulate the microbiome of seaweeds, providing a plausible mechanism for their ongoing loss, and encourage more research into the effects of human impacts on crucial but yet largely unstudied host-microbiome relationships in different aquatic and terrestrial species.

Evenness, biodiversity, and ecosystem function of intertidal communities along the Italian coasts: Experimental short-term response to ambient and extreme air temperatures.

Biodiversity can promote ecosystem functioning in both terrestrial and marine environments, emphasizing the necessity of biodiversity conservation in order to preserve critical ecosystem functions and associated services. However, the role of biodiversity in buffering ecosystem functioning under extreme events caused by climate change remains a major scientific issue, especially for intertidal systems experiencing stressors from both terrestrial and marine drivers. We performed a regional-scale field experiment along the Italian coast to investigate the response of unmanipulated intertidal communities (by using a natural biodiversity gradient) to low tide aerial exposure to both ambient and short-term extreme temperatures. We specifically investigated the relationship between Biodiversity and Ecosystem Functioning (BEF) using different biodiversity indexes (species richness, functional diversity and evenness) and the response of the intertidal communities' ecosystem functioning (community respiration rates). Furthermore, we investigated which other environmental variables could influence the BEF relationship. We show that evenness explained a greater variation in intertidal community ecosystem functioning under both temperature conditions. Species richness (the most often used diversity metric in BEF research) was unrelated to ecosystem functioning, while functional diversity was significantly related to respiration under ambient but not extreme temperatures. We highlight the importance of the short-term thermal history of the communities (measured as body temperature) in the BEF relationship as it was consistently identified as the best predictor or response under both temperature conditions. However, Chlorophyll a in seawater and variation in sea surface temperature also contributed to the BEF relationship under ambient but not under extreme conditions, showing that short-duration climate-driven events can overcome local physiological adaptations. Our findings support the importance of the BEF relationship in intertidal communities, implying that systems with more diverse and homogeneous communities may be able to mitigate the effects of extreme temperatures.

The current treatment of hip arthroplasty revision: a systematic review of the literature.

INTRODUCTION: Acetabular revision surgery is the most challenging aspect in hip prosthetic. There is lack of consensus on the optimal method of reconstructing the acetabular defects. The aim of this systematic review is to take stock of the state of the art on the options available and highlight which type of construct is the most reliable in usual clinical practice. MATERIAL AND METHODS: The reporting of this systematic review was guided by the standards of the Preferred Reporting Items for Systematic Review and Meta-Analysis 2020 Statement.Electronic search of MEDLINE was performed from 1991 up to April 2021 to identify relevant studies for this review.  Discussion: various surgical techniques have been adopted and proposed to treat acetabular bone defects: cemented cups, large-sized non-cemented acetabular cups, higher positioned cups, reinforcement rings, cage, oblong cups, custom triflange implants, high porous metal cups and augments. Bone defect defines the type of components to be implanted and among those, outcomes are various depending on the study taken into account, the component used and the degree of initial bone defect. CONCLUSIONS: In acetabular revision surgerythe use of TM cups and augment is a valid option in presence of major bone loss and pelvic discontinuities. In clinical practice the use of TM components replaced rings, while the cup-cage implant replaced conventional cages. TM augments and cups can be considered as the most promising technique in the reconstruction of wide acetabular defects, while the use of cages can be considered as a valid option in the elderly population.

Insight on bacteria communities in outdoor bronze and marble artefacts in a changing environment.

Epilithic bacteria play a fundamental role in the conservation of cultural heritage (CH) materials. On stones, bacterial communities cause both degradation and bioprotection actions. Bronze biocorrosion in non-burial conditions is rarely studied. Only few studies have examined the relationship between bacteria communities and the chemical composition of patinas (surface degradation layers). A better comprehension of bacterial communities growing on our CH is fundamental not only to understand the related decay mechanisms but also to foresee possible shifts in their composition due to climate change. The present study aims at (1) characterizing bacterial communities on bronze and marble statues; (2) evaluating the differences in bacterial communities' composition and abundance occurring between different patina types on different statues; and (3) providing indications about a representative bacterial community which can be used in laboratory tests to better understand their influence on artefact decay. Chemical and biological characterization of different patinas were carried out by sampling bronze and marble statues in Bologna and Ravenna (Italy), using EDS/Raman spectroscopy and MinION-based 16SrRNA sequencing. Significant statistical differences were found in bacterial composition between marble and bronze statues, and among marble patinas in different statues and in the same statue. Marble surfaces showed high microbial diversity and were characterized mainly by Cyanobacteria, Proteobacteria and Deinococcus-Thermus. Bronze patinas showed low taxa diversity and were dominated by copper-resistant Proteobacteria. The copper biocidal effect is evident in greenish marble areas affected by the leaching of copper salts, where the bacterial community is absent. Here, Ca and Cu oxalates are present because of the biological reaction of living organisms to Cu ions, leading to metabolic product secretions, such as oxalic acid. Therefore, a better knowledge on the interaction between bacteria communities and patinas has been achieved.

Early failure of cement with loosening and dislocation of the femoral component in a unicompartmental knee replacement: a case report with microscopic assessment.

Background Medial unicompartimental knee artrhoplasty (UKA) is a valuable and well-known option in the treatment of medial osteoarthritis (OA). Early recovery and good results are usually reported. Failure mechanism include septic and mechanical loosening, bearing dislocation and lateral or patello-femoral joint OA evolution. The rare case of an atraumatic dislocation of the cemented femoral component of a UKA is presented together with a literature review and a microscopic analysis of the loosened component. Methods The case of a 60-years old man who suffered a UKA failure due to a complete loosening and migration of the cemented femoral component 5 months after its implantation is reported. A review of the literature pertaining early similar catastrophic failures is discussed. Furthermore a stereo-microscopic and scanning electronic microscopic evaluation of the femoral component was performed. Results A UKA-to-TKA revision was performed. Septic loosening was ruled out and one-year follow up showed patient satisfaction with good clinical and radiographic results. Few cases of complete dislocation of the UKA femoral component are reported in the literature. Macro- and microscopic evaluation showed an almost completely smooth surface at the cemented surface of the posterior condyle of the femoral component.   Conclusions Whilst mobile bearing dislocation is a well-known complication of UKA, few cases of this potentially catastrophic complication are reported in the literature. Early UKA failure with complete implant loosening may be determined by a suboptimal cementing technique with inadequate cement penetration into the trabecular bone. In the present case, the absence of cement penetration into the posterior condyle may be one of the reason of the component dislocation after standing up starting with the knee in a highly flexed position.

Outcomes in arthroscopic surgery and proposal rehabilitative treatment in femoral acetabular impingement syndrome.

Background and aim of the work Femural Acetabular impingement syndrome (FAIS) is a patologic condition that can lead to hip pain, functional limitation and stiffness. In the last few decades orthopedics and physiotherapists have improved both surgery and riabilitative treatment leading to a better and better treatment. The target of this paper il to verify the efficiency of an early and multimodal physiotherapic treatment after and arthroscopic surgery of the FAIS   Materials and Methods We performed arthroscopic treatment and rehabilitation on 19 patients with mean age of 37±8,3 years, 12 males and 7 females. Each patient has been evaluated preoperatively (T0), postoperatively after 6 week (T1) and after 3 months of follow up (T2), the assessment was carried out by: administration of the VAS and WOMAC score for pain and function and joint examination of active hip movement through an inertial sensor system.   Results VAS score shows a decrease of pain after 6 week (mean decrease was 36%) and after 3 months (mean decrease was 33%). WOMAC score shows an increase of the funcional performance of the hip after 3 weeks and after 3 months (in both phases the mean score increase of the 44%) At last, the analysis of the active movement and of the hip joint showed a generalized increase in all movements both 6 weeks and 3 months after surgery, in particular for flexion (with the knee flexed) and internal rotation movements. of the hip.   Conclusion The results of this study are in line with the current scientific literature and the protocol used represents a valid tool to complete the surgical treatment. The proposal of an early, intensive treatment combined with hydrokinesitherapy seems to be safe and effective, however further studies are needed (increasing the sample size) to investigate the results.

A Comprehensive Biomarker Analysis of Microsatellite Unstable/Mismatch Repair Deficient Colorectal Cancer Cohort Treated with Immunotherapy.

The search for immunotherapy biomarkers in Microsatellite Instability High/Deficient Mismatch Repair system (MSI-H/dMMR) metastatic colorectal cancer (mCRC) is an unmet need. Sixteen patients with mCRC and MSI-H/dMMR (determined by either immunohistochemistry or polymerase chain reaction) treated with PD-1/PD-L1 inhibitors at our institution were included. According to whether the progression-free survival with PD-1/PD-L1 inhibitors was longer than 6 months or shorter, patients were clustered into the IT-responder group (n: 9 patients) or IT-resistant group (n: 7 patients), respectively. In order to evaluate determinants of benefit with PD-1/PD-L1 inhibitors, we performed multimodal analysis including genomics (through NGS panel tumour-only with 431 genes) and the immune microenvironment (using CD3, CD8, FOXP3 and PD-L1 antibodies). The following mutations were more frequent in IT-resistant compared with IT-responder groups: B2M (4/7 versus 2/9), CTNNB1 (2/7 versus 0/9), and biallelic PTEN (3/7 versus 1/9). Biallelic ARID1A mutations were found exclusively in the IT-responder group (4/9 patients). Tumour mutational burden did not correlate with immunotherapy benefit, neither the rate of indels in homopolymeric regions. Of note, biallelic ARID1A mutated tumours had the highest immune infiltration and PD-L1 scores, contrary to tumours with CTNNB1 mutation. Immune microenvironment analysis showed higher densities of different T cell subpopulations and PD-L1 expression in IT-responders. Misdiagnosis of MSI-H/dMMR inferred by discordances between immunohistochemistry and polymerase chain reaction was only found in the IT-resistant population (3/7 patients). Biallelic ARID1A mutations and Wnt signalling activation through CTNNB1 mutation were associated with high and low T cell immune infiltrates, respectively, and deserve special attention as determinants of response to PD-1/PD-L1 inhibitors. The non-MSI-H phenotype in dMMR is associated with poor benefit to immunotherapy. Our results suggest that mechanisms of resistance to immunotherapy are multi-factorial.

Genetic evolution to tyrosine kinase inhibitory therapy in patients with EGFR-mutated non-small-cell lung cancer.

BACKGROUND: Tumour heterogeneity impacts the efficacy of metastatic cancer treatment even if actionable mutations are identified. Clinicians need to understand if assessing one lesion provides reliable information to drive a therapeutic decision in non-small-cell lung cancer (NSCLC) patients. METHODS: We analysed inter-tumour heterogeneity from five autopsied individuals with NSCLC-harbouring mutations in the epidermal growth factor receptor (EGFR), treated with EGFR tyrosine kinase inhibitors (TKIs). Through a comprehensive next-generation sequencing (NGS) oncopanel, and an EGFR panel for digital droplet PCR (ddPCR), we compared metastases within individuals, longitudinal biopsies from the same lesions and, whenever possible, the primary naive tumour. RESULTS: Analysis of 22 necropsies from five patients revealed homogeneity in pathogenic mutations and TKI-resistance mechanisms within each patient in four of them. In-depth analysis by whole-exome sequencing from patient 1 confirmed homogeneity in clonal mutations, but heterogeneity in passenger subclonal alterations. Different resistance mechanisms were detected depending on the patient and line of treatment. Three patients treated with a c-MET inhibitor in combination with TKI lost MET amplification upon progression. CONCLUSION: At a given point and under selective TKI pressure, a single metastasis biopsy in disseminated tumours from EGFR-mutated NSCLC patients could provide a reasonable assessment of actionable alterations useful for therapeutic decisions.

Are vascular grafts at risk of occlusion after total hip replacement? A systematic review of the literature.

Vascular complications during and after total hip replacement are relatively uncommon despite the close relationship between the involved structures. Previous surgical procedure of arterial reconstruction or substitution may be at higher risk of damage due to the modification in the anatomical landmarks or to the mechanical properties of the grafts different from native vessels. In literature few cases of graft occlusion or failure are reported during or soon after a total hip replacement. The aim of this review is to report them highlighting common features and risk factors.

Kinetic and Kinematic analysis of ACL reconstruction in association with lateral-extrarticular tenodesis of the knee in revision surgery: a pilot study.

Background Anterior cruciate ligament reconstructions are a very frequent surgery. The key role of the anterolateral ligament in the knee rotational stability has been undelighted in recent years. Extra-articular tenodesis in association with ACL reconstructions, serves to eliminate anterolateral rotatory instability. The aim of our study is to compare treated knees with the contralateral uninjured knee in those cases whom been treated with ACL arthroscopic reconstruction and lateral extra-articular tenodesis in revision surgery, and evaluate clinical results with kinematic and kinetic examinations methods. Materials and methods Sixteen patients (10 males and 6 females) with ages from 21 to 37 had been treated at the Orthopaedic Clinic of Udine for failure of previous ACL reconstruction. In all patients was performed ACL arthroscopic reconstruction with the association of a lateral extra-articular tenodesis (Coker-Arnold). We have decided to asses the patients at one year after surgery with GNRB arthrometer and Bioval inertial sensor system. Results All patients treated with arthroscopic ACL reconstruction and lateral extra-articular tenodesis have regained pre-injury sagittal knee stability and gait dynamics.

Treatment options for massive rotator cuff tears: a narrative review.

The treatment of massive rotator cuff tears poses a challenge to orthopedic surgeons. The prevalence of massive rotator cuff tears is 40% of all rotator cuff tears. Compared with smaller tears, massive rotator cuff tears are often complicated by structural failure and poor outcomes and present a higher rate of recurrent tearing after surgical repair. Several management options are available but the selection of the most appropriate treatment for each patient can be challenging. To achieve the best outcomes, the orthopedic surgeon should have a good understanding of the indications, the pathomechanics and the clinical outcomes of the various treatment modalities. Treatment options include non-operative management, arthroscopic debridement with a biceps tenotomy or tenodesis, complete or partial repair, patch augmentation, superior capsular reconstruction, muscle/tendon transfer and reverse total shoulder arthroplasty. The purpose of this article is to review treatment options and clinical outcomes for the management of massive rotator cuff tears.

Cementless reverse total shoulder arthroplasty in a patient affected by Osteogenesis Imperfecta: a case report and review of the literature.

BACKGROUND AND AIM OF THE WORK: Osteogenesis Imperfecta is a rare genetic condition. The use of total shoulder arthroplasty in these patients is very uncommon, with only two cases reported in the literature. This study describes a cementless reverse total shoulder arthroplasty (rTSA) for a multi-fragmented fracture in a patient affected by Osteogenesis Imperfecta (OI) type 1 and aims to review literature results of shoulder replacement in patients affected by this uncommon condition. METHODS: the case of a woman affected by OI type 1 treated with a cementless rTSA for a multi-fragmented proximal humerus facture is reported. Focusing on the fixation technique, a research of all articles regarding the orthopaedic treatments in patients affected by Osteogenesis Imperfecta was performed and compared to the techniques used in the unaffected population. RESULT: Our patient shows good results in terms of clinical and radiological outcomes at the short term follow up. Few studies treat the orthopaedic manifestation of this rare genetic condition and only two are focused on shoulder arthroplasty. Cement is the preferred method for fixation in both papers. CONCLUSION: Cementless reverse shoulder arthroplasty may be an option in patients affected by OI type 1, although literature seems to support cement as the fixation method of choice.

MYC Copy Number Detection in Clinical Samples Using a Digital DNA-Hybridization and Detection Method.

Clinical tumor specimens are routinely formalin-fixed and paraffin-embedded (FFPE) in Pathology departments worldwide. FFPE blocks are convenient, long-term stable, and easy to archive and manipulate. However, nucleic acids extracted from FFPE tissues generally show a high degree of fragmentation as well as chemical modifications, mainly due to the fixation process. Methods to determine copy number alterations (CNAs) from FFPE clinical samples have proven challenging, in the fact that they are low-plex, only able to profile single genes or gene clusters (such as in situ hybridization-based methods), and/or show a low degree of robustness with partially degraded samples (array-based, NGS-based) as well as being time-consuming, costly, and with limitations in resolution. The NanoString nCounter® System is a medium-plex, extremely FFPE-robust system, that overcomes several of the frequent issues when dealing with clinical samples. The technique is based on hybridization of molecular barcoded probes directly to FFPE-derived DNA, followed by single molecule imaging to detect hundreds of unique molecules in a single reaction without any amplification steps that might introduce undesired biases. Here we describe nCounter v2 Cancer Copy Number Assay, a robust and highly reproducible method for detecting the copy number status of 87 genes commonly amplified or deleted in cancer, including the MYC proto-oncogene.

Molecular profiling of long-term responders to immune checkpoint inhibitors in advanced non-small cell lung cancer.

Immunotherapy has transformed advanced non-small cell lung cancer (NSCLC) treatment strategies and has led to unprecedented long-lasting responses in some patients. However, the molecular determinants driving these long-term responses remain elusive. To address this issue, we performed an integrative analysis of genomic and transcriptomic features of long-term immune checkpoint inhibitors (ICIs)-associated responders. We assembled a cohort of 47 patients with NSCLC receiving ICIs that was enriched in long-term responders [>18 months of progression-free survival (PFS)]. We performed whole-exome sequencing from tumor samples, estimated the tumor mutational burden (TMB), and inferred the somatic copy number alterations (SCNAs). We also obtained gene transcription data for a subset of patients using Nanostring, which we used to assess the tumor immune infiltration status and PD-L1 expression. Our results indicate that there is an association between TMB and benefit to ICIs, which is driven by those patients with long-term response. Additionally, high SCNAs burden is associated with poor response and negatively correlates with the presence of several immune cell types (B cells, natural killers, regulatory T cells or effector CD8 T cells). Also, CD274 (PD-L1) expression is increased in patients with benefit, mainly in those with long-term response. In our cohort, combined assessment of TMB and SCNAs burden enabled identification of long-term responders (considering PFS and overall survival). Notably, the association between TMB, SCNAs burden, and PD-L1 expression with the outcomes of ICIs treatment was validated in two public datasets of ICI-treated patients with NSCLC. Thus, our data indicate that TMB is associated with long-term benefit following ICIs treatment in NSCLC and that TMB, SCNAs burden, and PD-L1 are complementary determinants of response to ICIs.

Evolving Landscape of Molecular Prescreening Strategies for Oncology Early Clinical Trials.

Most academic precision oncology programs have been designed to facilitate enrollment of patients in early clinical trials with matched targeted agents. Over the last decade, major changes were seen both in the targetable molecular alteration landscape and in drug development trends. In this article, we describe how the Vall d'Hebron Institute of Oncology molecular prescreening program adapted to a dynamic model of biomarker-drug codevelopment. We started with a tumor-agnostic hotspot mutation panel plus in situ hybridization and immunohistochemistry of selected markers and subsequently transitioned to tumor-specific amplicon-based next-generation sequencing (NGS) tests together with custom copy number, fusion, and outlier gene expression panels. All assays are optimized for archived formalin-fixed paraffin-embedded tumor tissues without matched germline sequencing. In parallel, biomarker-matched trials evolved from a scenario of few targets and large populations (such as PI3K inhibitors in PIK3CA mutants) to a complex situation with many targets and small populations (such as multiple targetable fusion events). Recruitment rates in clinical trials with mandatory biomarkers decreased over the last 3 years. Molecular tumor board meetings proved critical to guide oncologists on emerging biomarkers for clinical testing and interpretation of NGS results. The substantial increase of immunotherapy trials had a major impact in target prioritization and guided clinical implementation of new markers, such as tumor mutational burden, with larger exon-based NGS assays and gene expression signatures to capture microenvironment infiltration patterns. This new multiomics era of precision oncology is expected to increase the opportunities for early clinical trial matching.