Academic literature on the topic 'Trial pre-clinico'

Abstract Purpose: Studies based on data from routine clinical practice (real-world data, RWD) benefit from larger patient numbers and are more representative of patient diversity than clinical trial studies. When combined with comprehensive genomic profiling (CGP), RWD may uncover the impact of therapies and patient characteristics on tumor genomic landscape. Here we aimed at assessing the feasibility of using RWD to identify changes in the prevalence of genomic alterations upon treatment and proposed a methodology to address RWD inherent caveats. Experimental Design: To explore associations between tumor genomics and treatment chosen by physicians, we evaluated data from 5323 patients with metastatic hormone receptor-positive HER2-negative (HR+/HER2-) breast cancers from a nationwide real-world clinico-genomic database, originating from approximately 280 US cancer clinics (~800 sites of care). To perform our comparisons, we defined groups based on the therapy administered in the metastatic setting and the timing of the CGP relative to treatment. We used bootstrapping to estimate the significance of the effect and stratified analyses to assess the impact of potential confounders such as the site of the collected samples or disease history. Results: ESR1 alteration prevalence increased from 5.6% (CI: 2.8-8.9) pre-treatment to 21.4% (CI: 13.3-29.6) following aromatase inhibitor. Yet, it was significantly less than the prevalence following treatments including CDK4/6 inhibitors (CDK4/6i; 37.1% [CI: 27.8-46.4]; P=0.006). Further, exposure to CDK4/6i led to an increase in FGFR1 and TP53 alterations as well as genes of the cell cycle (FDR< 0.2). Overall, we found that more pathways were likely to be altered in a given tumor following AI+CDK4/6i than after AI alone (P=0.02). In particular, alterations of the MAPK pathway were not exclusive to ESR1 alterations in the post-AI+CDK4/6i group compared to AI only, suggesting that MAPK pathway alterations alone may not overcome CDK4/6i-based treatments. Differences following exposure to CDK4/6i were retained in samples taken after the second-line treatment. Stratified analyses confirmed that these results are independent of exposure to adjuvant therapy or treatment duration and showed that ESR1 mutations occurred in both primary and metastatic samples. Conclusions: Analysis of EHR-derived clinical data linked to CGP results from routine care can replicate associations previously observed in clinical trials and uncover unknown changes in tumor genomic landscape. Bootstrapping and stratified analysis reinforced our confidence in the results and thus allowed us to identify that CDK4/6i exposure led to a different – more altered – genomic landscape in HR+/HER2- breast cancer patients. This finding can inform design of clinical trials post-CDK4/6i and may help guide treatment choice for late stage patients. Thus, our work demonstrates the feasibility of leveraging real-world clinico-genomic data for translational research in oncology and leads the way for analyses including a more diverse patient population. Citation Format: Nayan Chaudhary, Ciara Mecalfe, Marc Hafner. Real-world clinico-genomic data reveal differences in genomic landscape associated with CDK4/6 inhibitors in HR+/HER2- breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P5-13-03.

514 Background: Neoadjuvant systemic therapy (NST) increases the frequency of breast-conserving therapy (BCT) in stage II-III breast cancer, but there is little data on how often it converts patients (pts) from BCT-ineligible (BCT-I) to BCT-eligible (BCT-E) and on the impact of other factors on surgical choices. We collected surgical assessment and management data from an international randomized trial of NST in triple-negative breast cancer (TNBC). Methods: Women with operable TNBC were randomized to veliparib (V) with carboplatin (C) and paclitaxel (P), placebo with C and P or placebo with P followed by doxorubicin and cyclophosphamide. The surgeons assessed BCT candidacy by clinico-radiographic criteria before and after NST; surgical management was at surgeon and patient discretion. We assessed interactions between BCT eligibility pre- and post-NST, germline BRCA mutation ( gBRCA) status, continent of treatment and achievement of pathologic complete response(pCR) and percentage of pts who underwent BCT versus mastectomy. Results: Pre- and post-NST surgical assessments were available for 604 pts who underwent surgery. BCT rates are listed in the Table. The BCT rate was 68% among pts deemed BCT-E after NST. pCR rates were identical between BCT-E pts who chose BCT (55%) vs. mastectomy (53%). Of 141 pts deemed BCT-I at baseline, 75 (53%) converted to BCT-E but only 42 (56%) of these opted for BCT. pCR rates were 49% in BCT-E converts vs. 36% in those remained BCT-I. gBRCA pts (n = 84) were less likely to choose BCT even if they were BCT-E. Pts treated in North America (NA) were less likely to choose BCT (55% vs. 80% for Europe and Asia P<0.0001) even among non- gBRCA considered BCT-E post-NST (61% vs. 85% P<0.0001). Conclusions: This largest prospective analysis of the impact of NST in TNBC demonstrates a conversion rate from BCT-I to BCT-E of 53%. BCT rates were lower in pts with gBRCA; the much higher mastectomy rate among BCT-E pts in NA merits investigation. Clinical trial information: NCT02032277. [Table: see text]

5076 Background: Treatment intensification with docetaxel or abiraterone improved survival for advanced prostate cancer starting androgen deprivation therapy (ADT) in the Systemic Therapy in Advancing or Metastatic Prostate Cancer: Evaluation of Drug Efficacy (STAMPEDE, NCT00268476) trial. However, survival and time-to-progression is highly variable on ADT, introducing the risk of unnecessary toxicity from additional treatments for some patients. Here we test the prognostic association of proliferation index using Ki67 scores in the control arm of the STAMPEDE population of high-risk localised (M0) and metastatic (M1) prostate cancer. Methods: Pre-ADT diagnostic needle biopsies were obtained from 517 men randomized in STAMPEDE arm A between 2006 and 2015. These were assessed for proliferation using an analytically optimised Ki67 immunohistochemistry assay. Ki67 was tested for associations with baseline clinico-pathological variables (Grade group, pre-ADT serum PSA and imaging metastatic burden) in univariable linear-regression models, and for associations with survival outcomes in multivariable Cox-regression models adjusted for these and additional confounding variables. Primary outcome measure was overall survival, secondary outcomes were prostate cancer-specific, failure-free, progression-free and metastatic progression-free survival. Results: Ki67 was available for 475 patients who received ADT only for at least 2 years ± radiotherapy. Of 202 M0, 74 were node positive. Of 273 M1, 116, 127 and 30 were respectively low, high and unknown radiological M1 volume. Ki67 score associated with higher Gleason (p=7.15x10-11) and presence of extra-pelvic metastases (p=1.41x10-8). Increasing Ki67 scores showed a strong linear association with poorer overall survival, with an estimated 2% increase in the hazard of death per percentage increase in the score (adjusted HR=1.02, 95% CI 1.01-1.02; p=1.04x10-5). There was also strong evidence that Ki67 associated positively with all secondary outcomes, including prostate cancer-specific survival (adjusted p=5.50x10-6) and metastatic progression-free survival (adjusted p=3.50x10-9). Conclusions: Ki67 immuno-score is strongly prognostic in clinically advanced prostate cancer independent of Gleason score and the other clinicopathological variables tested in this study. Ki67 is a clinically scalable assay that could improve selection for treatment intensification and provide a tool for screening patients most likely to benefit from further molecular investigation.

5021 Background: Men with advanced hormone-sensitive prostate cancer (HSPC) starting long term androgen deprivation therapy (ADT) follow a highly variable clinical course. Treatment intensification with docetaxel or AR targeted therapies improves outcomes but there is a risk of overtreatment, especially in non-metastatic (M0) or metastatic (M1) low volume disease. We established a framework for biomarker evaluation in the STAMPEDE trial. We aimed to evaluate the feasibility and clinical utility of assessing the burden of copy number (CN) aberrations in newly diagnosed advanced HSPC. We hypothesised that increased percentage genome altered (PGA) would associate with higher disease burden and worse prognosis. Methods: We implemented a scalable strategy using low coverage whole genome sequencing (lpWGS) of formalin fixed paraffin embedded (FFPE) diagnostic core biopsies from STAMPEDE participants randomised to the standard of care ADT arm, between 2005 and 2016. Tissue was retrieved from 136 trial sites. 315 cases were randomly selected, aiming for a biomarker population of 300, anticipating an assay failure rate ̃5%. We defined 40% as the minimum histopathologically determined tumor cellularity (TC) for inclusion. We performed a survival analysis investigating PGA at diagnosis as a continuous measure with fractional polynomial specification in Cox models adjusting for disease burden, Gleason grade, pre-ADT PSA (log-transformed), age at randomisation and TC. We pre-specified that all hypothesis tests required evidence at the 5% significance level to consider rejecting the null hypothesis. Results: We successfully CN profiled 300/315 cases. There were no significantly different baseline clinico-pathological features between the full trial comparison n = 3106 and final biomarker population n = 300, 290/300 cases were de novo presentations. PGA in the core with highest Gleason grade and TC was median 18% (range 0%-75%; n = 300). PGA was significantly higher in M1 (n = 169) compared to M0 (n = 131) cases (median: 21% vs 14%; p = 0.00006). 284/300 were subclassified by disease burden into M0 node negative and node positive, and M1 low and high volume. PGA was significantly associated with increased disease burden (p = 0.00002). Increased PGA was significantly and non-linearly associated with an increased hazard of failure-free survival (p = 0.004), progression-free survival (p = 0.002), metastatic progression-free survival (p = 0.003), overall survival (p = 0.045) and prostate cancer-specific survival (p = 0.011). Conclusions: Evaluation of the burden of CN aberrations in archival, poor quality FFPE diagnostic tissue from men randomised in the STAMPEDE trial is feasible using lpWGS and has potential clinical utility to identify better prognosis advanced HSPC patients, who may not require treatment intensification.

147 Background: The CROSS trial established the role of neoadjuvant radiation in the treatment of esophageal adenocarcinoma (EAC). While response to radiation is an important factor in predicting long-term outcomes, the vast majority of patients succumb to systemic disease. The purpose of this study is to assess predictors of survival in patients with EAC following radiation therapy. Methods: All patients who underwent resection after radiation therapy for EAC at a single institution were retrospectively identified from January 2004 to December 2014. Patients who died within 30 days of surgery were excluded. Cox-proportional hazard analyses were performed to identify clinico-pathological factors associated with survival after surgery. Results: In the time period, 334 patients underwent esophagectomy for EAC. Univariable/multivariable analyses are shown in the table. The presence of a pathologic complete response (pCR) did not correlate to survival. The most important factors in predicting outcome were pre-operative albumin and initial lymph node stage by endoscopic ultrasound (EUS). Pre-treatment N0 patients had better survival than N1 patients (median survival 37.2 vs. 16.3 months, P < 0.0001). Patients who remained N0 after radiation had much better outcomes than those that either developed N1 disease after radiation or were initially staged as N1 (stayed N0, N = 126, median survival 52.0 months; N1→N0, N = 85, median survival 22.9 months; N0→N1, N = 30, median survival 15.3 months; persistent N1, N = 44, median survival 11.4 months; P < 0.0001). Conclusions: Pathologic response to radiation does not predict outcomes for patients with EAC. Patients with node positive EAC have poor outcomes even after neoadjuvant radiation therapy. These patients are at an increased risk of distant disease and should be offered additional systemic therapies prior to surgical resection. [Table: see text]

e20507 Background: It is reported that about 20% of patients with resected NSCLC adenocarcinoma harbor an EGFR driver mutation in the United States. Up to the recent approval of osimertinib in the adjuvant setting for resected EGFR + NSCLC based on the ADAURA trial, routine molecular profiling of early-stage lung cancer had not been standard of care. We hypothesize that there is a significant proportion of patients with resected adenocarcinoma with unknown EGFR status who could benefit from treatment that are missed with our current testing practices. Methods: We performed a retrospective analysis of Stage IB-IIIA lung adenocarcinomas resected at the University of Miami from 2014 to 2019. Eligible patients were identified from the Cancer Registry and information on EGFR mutation testing and treatment was obtained from chart review. We evaluated the prevalence of EGFR mutation testing in this population and outcomes based on EGFR mutation status. Disease free survival (DFS) and clinico-pathologic characteristics were evaluated. We estimated the number of patients that would have been eligible for EGFR testing and adjuvant osimertinib therapy in the pre-ADAURA era in our patient cohort. Results: A total of 120 patients had resected stage IB-IIIA adenocarcinoma during this five-year period (Stage IB 42.5%; Stage IIA 13.3%; Stage IIB 25%; Stage IIIA 19.2%) with a median age of 66 years. Most were females (59%), NHWs (51.5%), Hispanics (46.9%), and former smokers (66.7%). Out of patients with Stage IB-IIIA NSCLC with adenocarcinoma, 42.5% completed recommended adjuvant platinum-based chemotherapy. Only 40% of patients were referred for EGFR testing during this study period. The prevalence of EGFR mutations in this population was 10.8% (13 /120), but 59% of cases had no available EGFR testing. The most prevalent mutation was L858R (53.8%) followed by exon-19 deletions (30.8%). A total of 6 patients received an EGFR TKI therapy during the follow up period (2 in the adjuvant setting). With a median follow up of 12 mos, the rate of recurrence by stage was: Stage IB (3.9%); Stage IIB (10%); Stage IIIA (13%). Median time to disease progression or death was 13 months in this subgroup. There was no difference in disease free survival for patients with EGFR testing and those without results available in this short follow up period. Conclusions: Based on this retrospective review, up to 60% of patients with early-stage NSCLC with non-squamous histology have no available EGFR testing in the pre-ADAURA era. Of the anticipated 20% of patients with expected EGFR mutations based on historical controls, we have only identified half of patients that would have been eligible for adjuvant osimertinib. This study establishes the importance of upfront EGFR mutation testing in all NSCLC patients, not only to prognosticate, but also to identify the subset of patients who could benefit from adjuvant EGFR therapy.

e20010 Background: Isatuximab and daratumumab target the CD38 transmembrane glycoprotein on MM cells. IKd and DKd regimens have shown reductions of HR=0.53 (95%CI 0.32-0.89) and HR=0.63 (95%CI 0.46-0.85) resp. in progression or death risk compared to Kd in RRMM. In the absence of a direct IKd vs DKd trial, we performed an indirect treatment comparison on progression free survival (PFS) to enable cost-effectiveness analyses. Methods: A 3-state (pre-progression, progression, death) partitioned survival model was specified. NMA-adjusted transition probabilities were estimated from fitted exponential functions (time horizon of 6 and 12 m; cycle length 28 days). Inputs included the Wholesale Acquisition Cost of IKd, DKd, and premedications; cost of medication administration; and cost of adverse event management. Utility inputs for pre-progression (0.65) and progression (0.61) were per literature. Costs and utilities were discounted at 3.5%/y. A payer perspective was adopted. Life years (LY), quality adjusted LY (QALY), and incremental cost-effectiveness (ICER) and cost-utility ratios (ICUR) were estimated in base case (BCA) and probabilistic sensitivity analyses (PSA). Cost-effectiveness acceptability curves (CEAC) were generated. Results: As detailed in the Table, 6m of IKd treatment was associated with incremental gains of 0.01 (PSA 0.01) LYs but no gains in QALYs at cost savings of $24,188 ($23,762), yielding a dominant ICER of $ -2,418,800 ($-2,376,200) per LYg (ICUR not estimable). Further, 12m of IKd treatment was associated with incremental gains of 0.04 (PSA 0.04) LYs (or 0.48m) and 0.02 (0.03) QALYs at incremental cost of $1,585 ($2,239), yielding ICER of $39,625 ($55,975) per LYg and ICUR of $79,250 ($74,633) per QALYg. Per CEAC, IKd is the dominated strategy in the 6m model and had probability of 50% of being cost-effective at WTP of $100,000 in the 12m model. Conclusions: Clinically, compared to DKd, IKd is associated with slight incremental gains in LYs of 0.12m over 6m and 0.48m over 1y. The 6m clinical gain comes with cost savings of approximately $24,000 or about 15% of IKd therapy, while the 12m gain requires a minimal cost commitment of around $2,000 or 0.6% of DKd treatment. These findings imply a clinico-economic benefit of isatuximab compared to daratumumab containing regimens in RRMM. [Table: see text]

BackgroundMethotrexate (MTX) is the gold standard, first-line therapy for rheumatoid arthritis (RA). However, not all patients respond to MTX, and the predictors of its response or non-response have not yet been reliably identified. Identification of these predictors will facilitate personalized therapeutic choices, and improve patient outcomes.ObjectivesTo identify the clinico-laboratory predictors of response to MTX monotherapy in active RA.MethodsThis study included patients with active RA (SJC≥2 and TJC≥4) aged 18-55 years, with disease duration <5 years, who were not receiving DMARDs (except HCQ and low-dose prednisolone) and had been enrolled in the multicentre, parallel group RCT comparing two different MTX escalation strategies in RA (MEIRA)1. All these patients received MTX monotherapy which was started at 15 mg/week, escalated to 25 mg/week by 4-8 weeks, and continued till 16 weeks. MTX response was defined as EULAR good or moderate response (based on DAS28-CRP-3v) at 16 weeks. Stepwise, multivariable logistic regression was done using key demographic (age, gender, BMI, comorbidities), clinical (disease duration, DAS28, HAQ), and laboratory parameters (RF, anti-CCP, ESR, CRP, RBC MTX-polyglutamates 1-4, IL-6, MMP-3) as independent variables to identify predictors of MTX response. A two-tailed p-value <0.05 was used for defining statistical significance. (Trial Reg: CTRI/2018/12/016549)ResultsOut of a total of 178 included patients [84% females, mean age 40 (9) years, mean DAS28-CRP=5.4 (1.1)], 113 (63.5%) were classified as MTX responders at 16 weeks. Age (OR=0.95, p=0.01), BMI (OR=1.12, p=0.006), and RF (OR=0.34, p=0.045) were found to be independent predictors of MTX response on multivariable analysis (Table 1). On sensitivity analysis with DAS28-ESR-based EULAR response, age (OR=0.94, p=0.003) and RF (OR=0.42, p=0.059) were replicated as independent predictors of MTX response, in addition to pre-treatment swollen joint count (OR=0.94, p=0.05).Table 1.Results of multivariable logistic regression analysis for prediction of response (as defined by DAS28-CRP-based EULAR good or moderate response) to methotrexate monotherapy in RAVariableOR (unadjusted)Unadjusted p-valueOR (adjusted)Adjusted p-valueAge0.97 (0.93-1.002)0.060.95 (0.91-0.99)0.01Male sex0.78 (0.35-1.76)0.55-BMI1.1 (1.02-1.19)0.011.12 (1.03-1.22)0.006Presence of comorbidities0.67 (0.31-1.44)0.31-Disease duration0.98 (0.79-1.22)0.87-Baseline DAS281.1 (0.81-1.49)0.54-Baseline HAQ1.04 (0.66-1.64)0.86-Baseline TJC1.01 (0.96-1.05)0.72-Baseline SJC0.97 (0.91-1.02)0.24-Baseline ESR1.01 (1.00-1.02)0.27-Baseline CRP1.00 (0.99-1.01)0.85-RF positive0.31 (0.11-0.85)0.020.34 (0.12-0.98)0.045Anti-CCP positive0.73 (0.27-1.99)0.54-MTX PG1 (16 weeks)0.99 (0.94-1.04)0.69-MTX PG2 (16 weeks)0.98 (0.95-1.02)0.37-MTX PG3 (16 weeks)0.99 (0.96-1.02)0.43-MTX PG4 (16 weeks)0.99 (0.95-1.03)0.62-Serum IL-6 (baseline)0.98 (0.95-1.02)0.33-Serum MMP-3 (baseline)1.00 (1.00-1.00)0.48-BMI= Body Mass Index, CCP= Cyclic Citrullinated Peptides, CRP= C-reactive protein, DAS= Disease Activity Score, ESR= Erythrocyte Sedimentation Rate, HAQ= Health Assessment Questionnaire, MTX= Methotrexate, PG= polyglutamate, OR=Odds Ratio, RF=Rheumatoid Factor, SJC= Swollen Joint Count, TJC= Tender Joint CountNote: Only variables with p-value <0.2 on univariable analysis were included in the multivariable analysis.ConclusionYounger age, RF negativity, higher BMI, and lower pre-treatment swollen joint count are potential predictors of response to MTX monotherapy in RA.References[1]Jain S, Dhir V, Aggarwal A, et al. Comparison of two dose escalation strategies of methotrexate in active rheumatoid arthritis: a multicentre, parallel group, randomised controlled trial. Ann Rheum Dis. 2021;80(11):1376-1384.Disclosure of InterestsNone declared.

Abstract PTCL are a heterogeneous group of uncommon lymphoid malignancies. Geographical differences have been reported, the NK/T-cell-derived nasal type being more common in Asia and the entheropathy-type more common in western countries. Except for the alk-positive anaplastic large cell lymphoma (ALCL) subtype, PTCL have generally a poorer outcome than their B-cell counterparts when treated with conventional therapeutic strategies. In recent years, as a result of an improved biologic understanding and definition of PTCL entities, an increasing number of PTCL-specific clinical trials have been initiated. The purpose of this analysis was to describe epidemiological and clinico-pathological features of the major subtypes in PTCL, as they occur in an unselected western population, in order to provide population-based data that may be useful for the design of future PTCL-specific studies. Although the LYFO registry was initiated in 1983, the present analysis only includes patients diagnosed in the 15-year period from 1990 (when immunhistochemistry was routinely applied to all biopsy specimen) to 2004. Within this 15 yr period, 485 PTCL cases were diagnosed. They had an age range of 16–92 yrs and a male to female ratio of 1.4 (59% male and 41% female cases). The most frequent histological subtypes were PTCL unspecified (PTCLu) (44%), non-cutaneous ALCL (alk-status not available) (35%) and angioimmunoblastic (AIL) (17%). The incidence trend for PTCL, taken as one group, did not show significant changes over the 15 years observation period. Approximately two thirds of the patients (65%) had disseminated disease (stage III–IV) at diagnosis, while half of the patients (49%) presented with B-symptoms. AIL had a higher frequency of cases with disseminated disease (93%, p<0.05), mainly due to bone marrow involvement, while ALCL had a significantly higher frequency of localized cases (46%, p<0.05). Surprisingly, the majority of PTCL patients (69%) were registered at diagnosis with a good performance score (WHO) of 0–1, with figures for PTCLu and ALCL of 65% and 69%, respectively, and slightly lower (52%, p>0.05) for AIL. All three major subtypes had predominantly nodal disease at presentation (PTCLu 56%, ALCL 57%, AIL 61%). Pre-therapeutic s-LDH was elevated in 40 % of all PTCL patients (PTCLu 44%, ALCL 39%). AIL cases presented more often with elevated s-LDH (61% of the cases, p<0.05). In line with these findings, AIL also had a higher frequency of cases with IPI ≥2, which translated into an inferior 5-yr overall survival value (25%, as opposed to 33% and 39% for PTCLu and ALCL, respectively). These survival values represent the comparative background on which the potential impact of the new PTCL trial program of our nordic collaborative group will be evaluated.

1024 Background: Using samples from TAMRAD study (Treilleux, Ann Oncol, 2015), we previously reported that p4EBP1, a downstream protein of mTOR, was associated with higher benefit to everolimus (eve). SAFIRTOR study was designed to validate clinical utility of this biomarker. Methods: Patients (pts) with ER+, HER2 negative, AI resistant MBC were prospectively included (NCT02444390). All pts had a biopsy of a metastatic site and were then treated with standard eve + exemestane (exe) combination. The primary end point was to validate that p4EBP1 expression is associated with longer PFS in patients treated with eve. 120 evaluable pts were needed for the pre planed statistical analysis. All samples were collected and processed in a standardized procedure in order to allow phophoproteins IHC staining. In addition to p4EBP1, we explored prognostic value of pS6K, pAkt, PTEN and LKB1, together with genomic alterations assessed by NGS and CGH arrays. Results: 150 pts were included, 30 pts had no adequate sample, and further 13 had missing clinical data, 107 were evaluable for primary objective. Median age was 62, they had previously progressed on AI treatment, either in the adjuvant (22 pts) or the metastatic setting (83 pts). 20 were considered as primary hormone resistant, 87 as secondary resistant. The median Allread score for p4EBP1 was 5.5 (range: 0-6.5). Analysis of the primary endpoint showed that p4EBP1 staining above the median is associated with a longer PFS on eve+exe. (median PFS: 9.3 months, 95CI 6.3-13.1 for high p4EBP1 versus 5.8 months, 95CI 3.7-7.8 for low p4EBP1, p = 0.02). Prognostic value of high pEBP1 remained significant when assessed in a multivariate analysis along classical clinico-biological prognostic factors for MBC (HR 0.57, 95%CI 0.38-0.88, p = 0.01). In this AI resistant population, the tumor of 42 (46%), 33 (35%) and 5 (5.3%) pts carried an activating mutation for ESR1, PIK3CA and AKT1, respectively. None of these mutational statuses were correlated to outcome. Conclusions: This prospective study validates p4EBP1 expression analysis to select patients most likely to benefit from everolimus + exemestane. Clinical trial information: NCT02444390.

L’ictus rappresenta la seconda causa di morte a livello globale e la prima causa di disabilità negli anziani. A causa di alcuni limiti di impiego, le principali terapie disponibili, volte a ripristinare il flusso sanguigno attraverso fibrinolisi e/o trombectomia meccanica, possono essere usate solo nel 60% dei pazienti con eziologia ischemica. Per tale motivo, la ricerca preclinica cerca di identificare target terapeutici utili per definire nuovi trattamenti ed estendere l’accesso alle cure. Nessuna nuova strategia terapeutica identificata negli studi preclinici ha però confermato la sua efficacia nei trial clinici. I problemi di traslazionalità dipendono da alcuni limiti degli studi preclinici, come l’uso di un ridotto numero di animali, che non rappresentano le condizioni reali dei pazienti o la mancata replicazione dei risultati ottenuti dai singoli laboratori, introducendo così bias metodologici, tra cui la mancanza di protocolli armonizzati. Il presente studio origina dalla necessità di migliorare il valore traslazionale della ricerca preclinica nell’ictus ischemico attraverso la definizione di un multicentre preclinical randomised controlled trial (pRCT), il progetto TRICS, che ha l’obiettivo di valutare l’azione neuroprotettiva del remote ischemic conditioning (RIC) dopo l’evento ischemico. Perché il RIC raggiunga una neuroprotezione significativa, l’outcome primario è che induca un miglioramento dei deficit sensorimotori a 48 ore dall’evento ischemico, calcolati attraverso il De Simoni neuroscore. Lo score valuta i deficit generali, che indicano lo stato di salute dell’animale e i deficit focali selettivamente associati alle conseguenze neurologiche dell’ischemia. Per ridurre al minimo i bias tecnici, il nostro studio ha previsto una prima fase di armonizzazione della valutazione comportamentale tra i centri, mediante analisi della concordanza tra i rater che eseguivano la misura, definita come interrater agreement. Questo è stato calcolato come intraclass correlation coefficient, ICC=0, nessun accordo, a ICC=1, accordo perfetto. Ci siamo posti l’obiettivo di raggiungere un interrater agreement con un ICC≥ 0.60. Una buona concordanza tra i centri è infatti una condizione indispensabile per garantire che tutti applichino lo stesso metodo di valutazione, assicurando l’oggettività, la trasparenza e la riproducibilità dei risultati. Dopo un training sull’esecuzione del De Simoni neuroscore, ogni centro ha indotto l’ischemia cerebrale nell’animale mediante l’occlusione transiente dell’arteria cerebrale media (tMCAo). A 48 ore dall’operazione, tutti gli animali sono stati filmati durante l’esecuzione del test e i video sono stati inviati al centro coordinatore, che li ha randomizzati e inoltrati ai centri. Un rater per ogni centro ha eseguito in cieco il De Simoni neuroscore sui video e sulle valutazioni è stata eseguita l’analisi statistica. Il risultato ottenuto dall’interrater agreement è stato di ICC=0.50 (0.22-0.77) per i topi e ICC=0.49 (0.210.77) per i ratti; non siamo riusciti a raggiungere l’obiettivo prefissato. Questi risultati sono dipesi da alcuni errori nell’esecuzione del test e dall’attribuzione sbagliata dei punteggi. È stata organizzata una seconda fase di training per superare le differenze di valutazioni più evidenti e preparati nuovi animali ischemici con cui sostituire i video che presentavano errori. Il risultato ottenuto dall’interrater agreement del secondo trial è stato di ICC=0.64 (0.37 – 0.85) per i topi e ICC=0.69 (0.44 - 0.88) per i ratti, soddisfacendo quindi il criterio per iniziare la fase interventistica dello studio. La fase di armonizzazione rappresenta un nuovo ed originale approccio nella ricerca preclinica e dovrebbe rappresentare lo schema di lavoro di base per pianificare uno studio pRCT al fine di renderlo solido e predittivo.
In view of fostering transferability of pre-clinical data on the efficacy of remote ischemic conditioning (RIC) in acute ischemic stroke, we designed two multi-centre translational trials in mice and rats of both sexes. We defined to model ischaemic stroke by the transient occlusion of the middle cerebral artery (tMCAo). The improvement of sensorimotor deficits at 48h after tMCAo in RIC-treated animals was defined as the primary outcome. This work presents the harmonization phase relative to the evaluation of sensorimotor deficits by De Simoni neuroscore. Each centre performed different tMCAo durations - 30, 45, 60 min - allowing sufficient variability in the outcome. Animals were monitored post-surgery according to the ARRIVE and IMPROVE guidelines and data was registered into an electronic case report form on RedCap. All animals were video recorded during the neuroscore and the videos (n=11 per species) were distributed and evaluated blindly by raters at all centres. The interrater agreement of neuroscore was described using intraclass correlation coefficient (ICC), ranging between ICC=0 (equivalent to chance) and ICC=1 (perfect agreement), setting a target of ICC≥0.60 as satisfactory. We obtained moderate agreement for mice (ICC=0.50 [0.22-0.77]) and rats (ICC=0.49 [0.21-0.77]). Errors were identified in animal handling and test execution. We thus performed a second training followed by a new blind evaluation replacing the videos with poor experimental execution. The interrater agreement improved for mice (ICC=0.64 [0.37-0.85]) and rats (ICC=0.69 [0.44-0.88]). In conclusion, two dedicated training on the neuroscore allowed us to reach the agreement target for both species and thus next proceed with the interventional phase of the project.