Tumor Biomarkers Research Articles

Managing hemolysis in serum neuron-specific enolase measurements – an automated algorithm for routine practice

Neuron-specific enolase (NSE) derived from neurons and peripheral neuroendocrine cells is a biomarker for neuroendocrine tumors and for prognostication in comatose cardiac arrest survivors. However, as platelets and erythrocytes contain NSE, hemolysis causes falsely elevated NSE. We used native serum and hemolysate derived from the same patients to make serial dilutions, and subsequently measured NSE (mNSE) and hemolytic index (HI) in each dilution. An algorithm suitable for the laboratory information system was developed based on the mNSE, HI and the estimated gradient of hemolytic interference from 30 patients. We estimated the associated uncertainty of the corrected NSE (cNSE) results based on the observed range of the gradient and derived an equation for cNSE for samples with limited hemolysis (i.e. 5 < HI ≤ 30): cNSE = mNSE − HI × (0.34 ± 0.23) µg/L. By semi-quantitatively grading the contribution from limited hemolysis, a texted result noting the hemolysis-associated degree of uncertainty can accompany the cNSE result. The major challenge of hemolysis when using serum NSE as a biomarker can be managed using an automated algorithm for correction of NSE results based on degree of hemolysis. However, laboratorians and clinicians should be aware of the limitations associated with in vivo hemolysis.

Predictive value of positive lymph node ratio in patients with locally advanced gastric remnant cancer.

e16119 Background: Traditional lymph node stage (N stage) has limitations in advanced gastric remnant cancer (GRC)patients; therefore, establishing a new predictive stage is necessary.To explore the predictive value of positive lymph node ratio (LNR) according to clinicopathological characteristics and prognosis of locally advanced GRC. Methods: Seventy-four patients who underwent radical gastrectomy and lymphadenectomy for locally advanced GRC were retrospectively reviewed. The relationship between LNR and clinicopathological characteristics was analyzed. The survival analysis was performed using Kaplan–Meier survival curves and Cox regression model. Results: Number of metastatic LNs, tumor diameter, depth of tumor invasion, Borrmann type, serum tumor biomarkers, and tumor–node–metastasis (TNM) stage were correlated with LNR stage and N stage. Univariate analysis revealed that the factors affecting survival included tumor diameter, anemia, serum tumor biomarkers, vascular or neural invasion, combined resection, LNR stage, N stage, and TNM stage (all P &lt; 0.05). The median survival time for those with LNR0, LNR1, LNR2 and LNR3 stage were 61, 31, 23 and 17 mo, respectively, and the differences were significant ( P = 0.000). Anemia, tumor biomarkers and LNR stage were independent prognostic factors for survival in multivariable analysis (all P &lt; 0.05). Conclusions: The new LNR stage is uniquely based on number of metastatic LNs, with significant prognostic value for locally advanced GRC, and could better differentiate overall survival, compared with N stage.

Multiplexed electrochemical nucleic acid sensor based on visible light-mediated metal-free thiol-yne click reaction for simultaneous detection of different nucleic acid targets

Simultaneous detection of multiple tumor biomarkers with a simple and low-cost assay is crucial for early cancer detection and diagnosis. Herein, we presented a low-cost and simple assay for multiplexed detection of tumor biomarkers using a spatially separated electrodes strategy. The sensor is fabricated based on a metal-free thiol-yne click reaction, which is mediated by visible light, on commercially available indium tin oxide (ITO) electrodes. Four biomarkers, including p53 DNA, Brca2 DNA, K-ras DNA, and MicroRNA-204 RNA, were used as model analytes, and the corresponding oligonucleotide probes were modified on the desired electrode units sequentially with 530 nm irradiation light in the presence of photosensitizer Eosin Y. By this visible light-mediated coupling reaction, oligonucleotide probe densities of up to 9.2 ± 0.7 × 1010 molecules/cm2 were readily obtained on the ITO electrode surface. The proposed multiplexed E-NA sensor could detect four different nucleic acid targets concurrently without crosstalk among adjacent electrodes and was also successfully applied for detecting targets in a 20% fetal calf serum sample. The detection limits for p53 DNA, Brca2 DNA, K-ras DNA, and MicroRNA-204 RNA were 0.72 nM, 0.97 nM, 2.15 nM, and 1.73 nM, respectively. The developed approach not only has a great potential for developing cost-effective biosensors on affordable substrates for nucleic acid target detection, but also be easily extended to detect other targets by modifying the specific oligonucleotide probes anchored on the electrode.

Clinical nSights: A software platform to accelerate real world oncology analyses.

e23316 Background: Real world data analyses are critical to assess drug safety and effectiveness, including in patient populations underrepresented in clinical trials. Significant barriers to performing such analyses remain, including: (1) incomplete access to electronic health record (EHR) data due to patient privacy concerns, (2) lack of data harmonization within and across EHRs, (3) requirements for time- and labor-intensive data curation efforts, and (4) necessity of coding or data science expertise given the absence of user-friendly platforms. We present Clinical nSights, a software platform that addresses these challenges to enable researchers and clinicians to perform rapid, robust, and scalable real world oncology analyses. Methods: With input EHR data from multiple health systems, we developed intuitive user interfaces and a scalable researcher friendly programming environment for real world clinical analytics. We used (1) nCognito, a previously described HIPAA-compliant de-identification algorithm, to de-identify EHRs across data modalities; (2) the combination of a de novo knowledge graph, vector and distribution similarity metrics, and manual review to harmonize data types including but not limited to laboratory tests, disease codes, and drugs; and (3) a series of Bidirectional Encoder Representations from Transformers (BERT) models to perform augmented curation of variables of interest (e.g., cancer staging, tumor biomarkers) from unstructured clinical records. We developed a user interface for users to create and analyze defined patient cohorts. Results: Clinical nSights facilitates intuitive cohort creation via flexible specification of inclusion and exclusion criteria, including options to perform temporal linking between criteria and propensity score matching. The ability to specify criteria documented in de-identified clinical notes expands the repertoire of available features for cohort creation beyond those captured in structured EHR fields, including but not limited to cancer stage and tumor biomarker status (e.g., HER2 status, microsatellite instability). Data harmonization enables scalable longitudinal analyses of laboratory test data across cancer types, including changes in measurements in the years leading up to and following initial cancer diagnosis. Augmented curation improves the coverage of phenotypes including immune-related toxicities of checkpoint inhibitors and symptoms experienced by patients prior to a cancer diagnosis. Using the no-code interface, users can easily quantify and visualize outcomes such as overall survival and hospitalization rates. Conclusions: Clinical nSights addresses important challenges in the collection, curation, and analysis of real world oncology data. By enabling a broad range of users to rapidly generate and test hypotheses, this platform has the potential to significantly improve care delivery and patient outcomes.

Neoadjuvant immunotherapy in sarcomatoid mesothelioma (Alliance A082101).

TPS8125 Background: Sarcomatoid mesothelioma is the most aggressive form of pleural mesothelioma and is associated with the worst prognosis of the histologic variants of this disease. In the Checkmate 743 clinical trial the patients with sarcomatoid mesothelioma who received ipilimumab and nivolumab had survival outcomes that were similar to those of patients with epithelioid mesothelioma. In comparison, the patients with sarcomatoid mesothelioma who received chemotherapy had the worst outcomes in this trial, highlighting the known limited efficacy of chemotherapy against this histologic variant. Checkmate 743 demonstrated that ipilimumab and nivolumab is a new standard of care for non-epithelioid mesothelioma. Surgery typically has not been offered to this group of patients given their historically poor outcomes. With the significant survival gains seen patients with sarcomatoid mesothelioma treated with immunotherapy, we hypothesized that surgery may extend the benefits seen with immunotherapy. Methods: Alliance for Clinical Trials in Oncology A082101 is a prospective, phase 2 nonrandomized clinical trial for patients with sarcomatoid mesothelioma. The co-primary objectives are to determine the percentage of patients with potentially resectable sarcomatoid mesothelioma able to proceed with surgery after neoadjuvant ipilimumab and nivolumab, and the progression-free survival (PFS) at 12 months (12-month PFS) after the initiation of ipilimumab and nivolumab. For sample size determination, we assumed that if the true rate of surgery is 75% or greater, it would indicate that neoadjuvant immunotherapy is feasible to be given prior to surgery. On the other hand, if the true rate of surgery is 50% or less it would indicate that neoadjuvant immunotherapy is not worthy of further investigation. Twenty-six (26) eligible patients will be needed to receive neoadjuvant immunotherapy. If 16 or fewer of the 26 eligible patients proceed to surgery, it will be concluded that the experimental therapy is not worthy of further investigation. Otherwise, it will be concluded that the experimental therapy is worthy of further investigation. Safety monitoring will be done through a 6-week run-in for the first 10 patients to assess for pre-operative and post-operative toxicities and complications. Sequential boundaries will be used to monitor severe toxicity and complication rates. Accrual will be halted if excessive numbers of severe toxicity and complications in pre-operative and post-operative phases are seen. The accrual and safety data will be summarized in reports for semiannual reviews by Alliance Data and Safety Monitoring Board. Tumor and blood-based biomarkers will be included as exploratory biomarkers. Support: U10CA180821, U10CA180882, U24 CA196171; https://acknowledgments.alliancefound.org . Clinical trial information: NCT05647265 .

Clinical and translational results from REVOLUTION cohorts A (nivolumab + ipilimumab + chemotherapy) and B (hydroxychloroquine + ipilimumab + chemotherapy) in patients with previously untreated metastatic pancreatic adenocarcinoma.

4137 Background: Metastatic pancreatic adenocarcinoma (mPDAC) is largely refractory to treatment, including immunotherapy. However, recent developments in chemoimmunotherapy combinations show promise. Herein, we report results from the first 2 cohorts of REVOLUTION, an ongoing, adaptive platform study designed to assess the safety and antitumor activity of novel chemoimmunotherapy combinations in patients (pts) with untreated mPDAC. Methods: REVOLUTION (NCT04787991) is an open-label, non-randomized, exploratory platform trial. Pts were assigned by investigator choice to one of the enrolling cohorts. All pts in Cohorts A and B received 1000 mg/m2 gemcitabine, 125 mg/m2 nab-paclitaxel, and 2 doses of 1 mg/kg ipilimumab. In addition, pts received 360 mg nivolumab Q3W in Cohort A or 600 mg hydroxychloroquine BID in Cohort B. Primary endpoint: safety. Secondary endpoints: ORR, DCR, DOR, PFS, and OS. Exploratory endpoints: pharmacodynamics and association of tumor and blood biomarkers with clinical activity. Each cohort utilized a Simon two-stage design: n=15 in Stage 1 with expansion to Stage 2 (an additional n=15) based on the totality of safety, efficacy and biomarker analyses. Results: Both cohorts enrolled 15 pts each. Median treatment duration was 6.5 and 5.6 months (mo), and median study duration was 19.1 and 12.7 mo for A and B, respectively. All pts experienced a treatment-related adverse event (TRAE). Grade 3-4 TRAEs occurred in 60% of A (most common: decreased neutrophil count [n=4]) and 53% of B (most common: anemia [n=5], decreased neutrophil count [n=4]). In B, 2 pts discontinued due to AEs, with none occurring in A. One grade 5 TRAE (multiple organ dysfunction syndrome, deemed related to chemotherapy) occurred in B; none in A. Table shows efficacy results. Cohort A revealed increased activated/proliferating CD4 T cells, CD8 T cells, Tregs, and soluble PD-1 in circulation. Cohort B showed increases in activated/proliferating CD4 T cells and signs of autophagy inhibition. Preliminary multi-omic analyses that aim to associate biomarkers with clinical response will be presented. Conclusions: REVOLUTION cohorts A and B demonstrated antitumor activity in pts with mPDAC. Cohort B treatment, which included hydroxychloroquine, incurred more early discontinuations. As designed, neither cohort will advance to Stage 2, highlighting the need for further tailored therapeutic approaches in mPDAC. Clinical trial information: NCT04787991 . [Table: see text]

Preliminary results of a phase 1, first-in-human, dose escalation study of the anti-CCR8 cytolytic antibody, CHS-114 (formerly SRF114) in patients with advanced solid tumors.

2664 Background: Depletion of intratumoral regulatory T cells (itTregs) represents an attractive therapeutic strategy to enhance antitumor responses. The chemokine receptor CCR8 is preferentially expressed on itTregs compared to peripheral Tregs and other immune cell types, and CCR8 expressing itTregs are highly immune suppressive. Our preclinical studies have demonstrated that treatment of a PD-1 resistant mouse tumor model with an anti-CCR8 monoclonal antibody (mAb) depletes itTregs, reduces tumor growth, and enhances anti-PD-1 antitumor activity. CHS-114 an afucosylated mAb that binds CCR8, depletes itTregs, and enhances toripalimab- (tori; anti-PD-1 mAb) mediated T cell activation. Here, we present preliminary results of the first-in-human phase 1 study of CHS-114. Methods: This phase 1, multicenter, single agent and combination (combo) dose escalation study (NCT05635643) of CHS-114 is enrolling patients (pts) ≥18 years of age with advanced solid tumors who progressed during or after standard therapy. CHS-114 dose escalation guided by the Bayesian optimal interval (BOIN) design will evaluate doses ranging from 5 to 1500 mg q3w. Five additional pts with advanced head and neck squamous cell carcinoma (HNSCC) will be enrolled at each of 2 selected dose levels (DLs) with required biopsies. Combo dose escalation will evaluate 2 DLs of CHS-114 combined with tori 240 mg q3w in pts with HNSCC using a standard 3+3 design. Primary objectives are to determine the recommended dose(s) for expansion of CHS-114 and the safety and tolerability of CHS-114 + tori. Secondary objectives are to evaluate the safety, tolerability, pharmacokinetics (PK), and preliminary antitumor activity of CHS-114 alone and in combination with tori. Tumor and immune biomarkers are being evaluated as exploratory endpoints. Results: As of 15Dec2023, 15 pts received CHS-114 at doses ranging from 5 to 700 mg q3w. No dose-limiting toxicities (DLTs) were reported. Treatment-related adverse events occurred in 33% of pts; all were CTCAE grade 1-2, with pyrexia (13%) being the most common. Preliminary PK analysis indicates that CHS-114 exposure increases with dose with an elimination half-life between 9-17 days across the dose range of 10-700 mg. Preliminary receptor occupancy analysis using an immune profiling assay in peripheral blood at 100, 300, and 700 mg DLs revealed minimal detection of CCR8+ Tregs and a modest concomitant decrease in overall Treg frequency at the end of cycle 1, consistent with strong receptor occupancy. Conclusions: CHS-114 has an acceptable safety profile to date in heavily pretreated pts at doses up to 700 mg. CHS-114 administration at 100 to 700 mg shows robust decreases in peripheral CCR8+ Tregs following the first dose (cycle 1). Dose escalation is ongoing. Clinical trial information: NCT05635643 .

Effect of a novel artificial intelligence (AI) –enabled multi-trial matching system on patient matching using real-world data.

e13501 Background: Currently clinical research teams at sites are manually matching patients to clinical trials, where each trial in our experience takes at least 25 minutes using highly trained dedicated resources. This limits cancer patients’ clinical trial options and enrollment rates. Moreover, the lack of NGS testing results may also negatively impact matching to clinical trials in targeted therapies. PCC, a consortium of 7 of the Top 20 Pharmaceutical companies, collaborated with Massive Bio to demonstrate how cancer patients’ matching rates can be increased if patients are prescreened to multiple trials simultaneously (i.e. multi-trial matching) instead of a single trial and NGS testing results are integrated into the matching process. Methods: Massive Bio has developed a first-in-class AI-enabled matching system to extract 180 structured clinical data from medical records using computer vision and natural language processing and a recommendation system for matching patients to parameterized inclusion/exclusion (I/E) criteria from over 14,000 actively recruiting interventional clinical trials.Our analysis included a sample of 5,707 cancer patients from the Massive Bio Synergy AI real-world database, corresponding to tumor types in 23 selected trials. We developed a decision-tree algorithm for retrospective matching analysis using modeled I/E criteria and compared these results to the theoretical matching rate (TMR) for each trial based on tumor type and biomarker prevalence, extent of disease at diagnosis and prior treatment history. Results: 690 patients matched to at least one trial were matched to 1.82 times more trials in multi-trial matching; 2.10 times more potential matches were obtained with hypothetical 100% NGS testing; and 99.9% matching time was reduced with AI enabled system; key findings are summarized (Table). Conclusions: This study confirms the underutilization of NGS technology and substantial benefits of AI and NGS in cancer trial matching. Their use resulted in a nearly two-fold increase in potential patient eligibility for trials, and an unprecedented reduction in manual effort, underscoring a significant advancement in the efficiency of the clinical trial matching process. A prospective country-wide analysis is under consideration. [Table: see text]

The NAVigate-HPV Registry: A comprehensive biomarker evidence base for HPV-driven cancers.

e15045 Background: Human papillomavirus (HPV) is an oncogenic driver of several human cancers, including those of the anal canal, uterine cervix, and head and neck (predominantly oropharynx). Despite advancements in our understanding of HPV-related cancers, there remains an unmet need for a comprehensive biomarker-driven registry to establish correlations between biomarker detectability and clinicopathologic features in patients diagnosed and treated for HPV-driven cancers. To address this need, the NAVigate-HPV Registry has been established to systematically collect and analyze integrated biomarker and clinical data from many U.S. cancer centers. Methods: NavDx (Naveris, Inc.) is a clinically validated blood-based assay detecting circulating cell-free tumor tissue modified viral (TTMV)-HPV DNA. The NAVigate-HPV Registry is a web-based data platform based on a centrally IRB approved protocol. Participating investigators will include providers that order NavDx tests in the routine clinical care of their patients and agree to the terms and conditions for use of the registry. Eligible subjects will have or have had an HPV-driven cancer (with an initial focus on HPV-driven oropharynx cancer with confirmatory tissue testing) and have received one or more NavDx blood tests during routine clinical care. Clinical data and TTMV-HPV DNA scores will be collected at enrollment and defined intervals over five years or until death. Results: In the initial phase, the registry has enlisted participation and established a steering committee composed of a representative from each of 14 institutions. These institutions are geographically diverse centers ranging from community hospitals to clinics and academic medical facilities. An IRB protocol governing the registry will maintain ethical standards, ensuring compliance and ethical conduct throughout the registry process. Data extraction methods have been previously established to evaluate an initial cohort of 543 patients (Hanna GJ, Roof SA, et. al 2023). Conclusions: Naveris and collaborators have together successfully launched the NAVigate-HPV registry with the aim of creating a robust database linking tumor biomarker results from NavDx with clinical outcomes data for HPV-related cancers. The NAVigate-HPV database is expected to encompass more than 1,000 patients within 1 year and grow to &gt; 5,000 patients within 5 years, serving as a powerful tool for precision cancer medicine in this patient cancer population. Establishing this registry will deepen insights into HPV-related cancers and advance diagnostic and therapeutic approaches to improve patient outcomes.

Use of liquid biomarker thymidine kinase activity (TKa) to predict outcome and progression in patients with metastatic breast cancer (MBC) in the GEICAM/2014-12 FLIPPER trial.

1028 Background: There is a need for more precise blood-based tumor biomarkers in MBC. TKa is a liquid proliferation biomarker providing prognostic, predictive and monitoring information in MBC. TKa can be measured in blood using the DiviTum TKa assay. Here we report the results of TKa in a trial of endocrine therapy (ET) fulvestrant (F) + the CDK4/6 inhibitor (CDK4/6i) Palbociclib (Pa) vs. F + Placebo (Pl), the GEICAM FLIPPER trial (NCT02690480), a multicenter, double-blind, randomized (1:1) study. F+Pa showed 1-year PFS rates of 83.5% vs. 71.9% (p=0.064) and better median (m)PFS (31.8 vs. 22.0 months, p=0.001) than F+Pl in first line treatment of HR+/HER2- MBC patients. F+Pa also significantly improved ORR and CBR. ET+CDK4/6i are currently the cornerstone of treatment for HR+/HER2- MBC. However, 10-30% of these patients display primary resistance to treatment. Early identification of patient subsets with different risk of progression during treatment with CDK4/6i may help tailor clinical management and disease monitoring. Methods: Sequential plasma was collected from 187 patients (pts) at baseline (BL) and at 12, 24, 36, 48 weeks (w) on treatment, and at progression. 910 samples were analyzed for TKa with the FDA approved and CE labelled DiviTum TKa assay (Biovica, Sweden) using DuA (DiviTum unit of Activity) as measuring unit. The cutoff for categorizing in high vs low is 250 DuA. The Kaplan-Meier method was used to estimate mPFS and mOS. Unadjusted and adjusted hazard ratio (HR) and odds ratio (OR) with 95% confidence interval (CI) were calculated with the Cox proportional hazards regression model and with logistic regression respectively, considering relevant confounders. Results: Low BL TKa predicted better PFS (30.5 months mPFS in low vs 19.8 in high, HR 1.62; 95% CI 1.11-2.35, p=0.011) and OS (HR 1.81; 95% CI 1.15-2.85, p=0.0099). Higher TKa at BL and at 12w was detected in pts that progressed before 12 months. Conversely, low BL TKa, maintained throughout week 12-48, were associated with objective response (HR 2.58; 95% CI 1.38-4.81, p=0.0029) and OS (HR 5.74; 95%CI 2.06-16, p=0.0008). At progression, pts on F+Pa tended to have higher TKa levels than F+Pl (246 vs. 138 DuA), reflecting faster growing tumors, not achieving statistical significance (p=0.0585). TKa monitoring and dynamics from BL and on treatment timepoints (week 12-48) show that the group of pts with low TK at BL that remain low during treatment is enriched with non-progressors and pts that progress after 12 months (p=0.0108). High TKa at BL predicted shorter OS in the F+Pa arm (HR 4.39; 95% CI 2.15-8.99, p&lt;0.0001). Conclusions: Measuring blood TKa levels can monitor and predict efficacy of F+/-Pa at BL and during therapy in MBC. TKa dynamics before and during therapy are associated with PFS and OS. Future studies will provide additional knowledge on the utility of TKa in MBC. Clinical trial information: NCT02690480 .

70P Tumor biomarkers contribute to the clinical management of the O-RADS MRI risk stratification system in epithelial ovarian tumors

70P Tumor biomarkers contribute to the clinical management of the O-RADS MRI risk stratification system in epithelial ovarian tumors

Procollagen C-protein enhancer 1 (PCPE-1) as a potential serum biomarker for osteosarcoma.

e23518 Background: Osteosarcoma (OS) is the most common bone malignancy in children and young adults. Treatment with perioperative systemic chemotherapy has improved patient outcome but a third of patients will eventually develop incurable metastatic disease. Currently, there are no biomarkers for monitoring patients’ response to neo-adjuvant (NA) chemotherapy or for patients under active surveillance following treatments. Procollagen C-Proteinase Enhancer-1 (PCPE-1) is an extra-cellular matrix glycoprotein which promotes the maturation of pro-collagen into collagen in various tissues. Elevated serum levels of PCPE-1 are found in bone diseases such as Paget’s disease and Osteoporosis and in patients with breast cancer with bone metastases. Serum PCPE-1 has not been described in patients with OS. We aimed to describe PCPE-1 serum levels in patients with OS compared with healthy volunteers, and across different time points in their treatment and disease course. Methods: We collected serum samples from the Israeli national biobank of 18 adult OS patients. Serum samples were also collected from 6 healthy individuals. Additionally, we obtained serial samples from 4 OS patients prior to and after initiating NA therapy. Quantification of serum PCPE-1 levels was performed using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. Results: We successfully analyzed serum PCPE-1 levels from all patients and healthy volunteers. Mean PCPE-1 levels of 553ng/ml (range 479-614ng/ml) were observed in OS patients, compared with 313ng/ml (285-339ng/ml) in healthy volunteers. Of 4 patients undergoing treatment who had serial samples, a decrease in serum levels was observed 2 responding patients, while an increase in PCPE-1 levels were observed in one patient after NA treatment who had no response to treatment at the time of surgery. Of notice, serum levels nearly doubled in a patient during active surveillance who experienced local disease recurrence (274ng/ml and 423ng/ml, respectively). Conclusions: This is the first study to show elevated PCPE-1 serum levels in patients with OS. Our study also demonstrates potential changes in PCPE-1 levels in response to treatment. We are expanding our study to include a pediatric population to investigate the role of PCPE-1 as a potential tumor biomarker for monitoring treatment and surveillance of OS.

Single point of entry to the European precision cancer medicine trial network PRIME-ROSE.

e23024 Background: Over the last decades, there has been a surge in the development and approval of targeted drugs and immunotherapies for treating cancer patients, as seen by the 757 % increase in approvals by the FDA for new cancer treatments since 2000 (1). This has significantly impacted cancer care and contributed to improving overall survival in various cancer subgroups. However, access to these new treatments is constrained by the market access strategy of the patent-owning company and available knowledge of treatment effects. Only a few treatments have received pan-cancer approval from EMA or FDA, and most drugs receive market authorization per indication. As a result, there is a widening access gap between patients with different cancer types (2). Methods: Exploring the effect of biomarker-driven treatments in new cancer subtypes requires the ability to find patients with rare biomarkers. PRIME-ROSE is a European precision medicine network comprising 11 ongoing or soon-to-start national DRUP-like clinical trials testing registered drugs outside their current label ( www.prime-rose.eu ). Patients with relevant tumor biomarkers are identified and treated with matched drugs available in each trial’s drug portfolio. The ambition is to swiftly and systematically evaluate the effectiveness of approved precision cancer medicines in new indications through pragmatic trial designs and with RWE control cohorts, ensuring expansion into all relevant patient groups to maximize societal benefit. This is particularly relevant for rare cancers, which are enriched in precision medicine trials (3). Results: In PRIME-ROSE, the trials now share and aggregate data to build evidence faster and more effectively impact patient care by addressing key challenges in precision cancer medicine implementation (increasing the recruitment area to 71 million inhabitants). This will significantly reduce the time for filling treatment cohorts and contribute to closing the indication/drug-specific knowledge gap. In fact, several pharmaceutical companies have already shown their interest in and commitment to participating in PRIME-ROSE, as it offers the unique advantage of entering the trials in the network simultaneously (single point of entry) and with a floating allocation of treatment slots between trials, increasing efficiency in finding patients with specific biomarkers to fill treatment cohorts. Conclusions: A unified entry point to the PRIME-ROSE network is feasible and can facilitate building the knowledge base faster for label expansion and/or country-specific approvals/ reimbursement. National multi-stakeholder ecosystems that include pragmatic, RWE-controlled DRUP-like clinical trials may advance precision medicine implementation.

Serum β-hCG as a Biomarker in Pancreatic Neuroendocrine Tumors: Rethinking Single-Analyte Approach.

Despite recent advances, neuroendocrine tumors (NETs) remain a challenging topic, due to their diversity and the lack of suitable biomarkers. Multianalyte assays and the shift to an omics-based approach improve on the conventional single-analyte strategy, albeit with their own drawbacks. We explored the potential of serum β-hCG as a biomarker for NETs and discussed its role in disease monitoring. We recruited 40 patients with non-functioning pancreatic NETs, all with liver metastases. Serum β-hCG concentrations were measured at 3-month intervals over 48 months. We performed a comparative and a repeated measures analysis of β-hCG depending on WHO grade (G1, G2), liver tumor burden (LTB; below 10%, 10-25%), and RECIST 1.1. (stable disease, progressive disease). Patients with progressive disease (p < 0.001), 10-25% LTB (p < 0.001) and WHO Grade 2 (p < 0.001) displayed higher β-hCG concentrations. Throughout the study, β-hCG concentrations consistently increased across the entire cohort. Delta β-hCG during the study period was greater in patients with 10-25% LTB (p < 0.001), progressive disease (p < 0.001), and G2 (p = 0.003). Serum β-hCG correlates with established indicators of malignancy and disease progression in metastatic NETs, supporting further studies as a monitoring and prognostic biomarker. Despite promising results from novel biomarkers, there is still a place for single-analyte assays in NETs.

Advancing MicroRNA Detection: Enhanced Biotin-Streptavidin Dual-Mode Phase Imaging Surface Plasmon Resonance Aptasensor.

MicroRNAs (miRNAs) are novel tumor biomarkers owing to their important physiological functions in cell communication and the progression of multiple diseases. Due to the small molecular weight, short sequence length, and low concentration levels of miRNA, miRNA detection presents substantial challenges, requiring the advancement of more refined and sensitive techniques. There is an urgent demand for the development of a rapid, user-friendly, and sensitive miRNA analysis method. Here, we developed an enhanced biotin-streptavidin dual-mode phase imaging surface plasmon resonance (PI-SPR) aptasensor for sensitive and rapid detection of miRNA. Initially, we evaluated the linear sensing range for miRNA detection across two distinct sensing modalities and investigated the physical factors that influence the sensing signal in the aptamer-miRNA interaction within the PI-SPR aptasensor. Then, an enhanced biotin-streptavidin amplification strategy was introduced in the PI-SPR aptasensor, which effectively reduced the nonspecific adsorption by 20% and improved the limit of detection by 548 times. Furthermore, we have produced three types of tumor marker chips, which utilize the rapid sensing mode (less than 2 min) of PI-SPR aptasensor to achieve simultaneous detection of multiple miRNA markers in the serum from clinical cancer patients. This work not only developed a new approach to detect miRNA in different application scenarios but also provided a new reference for the application of the biotin-streptavidin amplification system in the detection of other small biomolecules.

An Interesting Case Report of Medullary Thyroid Carcinoma Co-Exisiting with Hashimoto’s Thyroiditis

Medullary thyroid carcinoma (MTC) is a rare malignant tumor, which accounts for &lt;5% of all thyroid malignancies, derived from parafollicular C cells. Hashimoto’s thyroiditis, which is the most common cause of hypothyroidism is an autoimmune disease. Many studies have been done in association with Hashimoto’s thyroiditis, Papillary thyroid carcinoma and thyroid lymphomas. Very few cases of Hashimoto’s thyroiditis with coexisting medullary thyroid carcinoma have been reported, because of its rarity. MTC produces various tumor biomarkers mainly Calcitonin, Carcinoembryonic antigen (CEA) and Chromogranin A. For determining the severity of the condition, preoperative serum calcitonin and CEA values are crucial. Postoperatively, Serum levels of calcitonin and CEA can be used to assess for distant metastases and recurrence. Here we present a rare case of Medullary thyroid carcinoma coexisting with Hashimoto’s thyroiditis.

Changes in expression of breast cancer tumor biomarkers between primary tumors and corresponding metastatic sites: common patterns and relationships with survival.

In metastatic breast cancer, differences in expression patterns of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2) between the primary tumor (PT) and metastatic site (MET) have been reported. However, there is limited understanding of the relationship of tumor subtype discordance and overall survival (OS). We evaluated patterns of ER/PR/HER2 in PTs and corresponding METs and assessed the relationship between these patterns and OS. Patients diagnosed at our center with metastatic breast cancer (2011-2020) were included. ER/PR were stratified as < 1%/1-10%/ > 10% by immunohistochemistry and HER2 as positive/negative by immunohistochemistry/FISH. Tumor subtypes were classified as ER or PR + /HER2-, HER2+ , or triple-negative. Biomarker discordance data from PTs to METs were analyzed for expression patterns. OS was assessed. Of 254 patients, 41 (16.1%) had synchronous and 213 (83.9%) had metachronous METs. Category change of ER/PR/HER2 expression was observed in 56 (22.0%), 117 (40.5%), and 30 (11.8%) patients, respectively. Tumor subtype changed in 56 (22.0%) patients. We identified a difference between PT and MET from ER > 10% to ER < 1% (n = 28,16.2% p < 0.01); PR > 10% to PR < 1% (n = 54,48.2%, p < 0.001); PR > 10% to PR 1-10% (n = 18,16.1%, p < 0.001), and ER or PR+/HER2-to triple-negative (n = 19,13.0%, p = 0.03). In log-rank analysis, change from an ER or PR+/HER2-(5-year OS 88.6%) PT to a HER2+(67.5%) or triple-negative (54.6%) MET was associated with decreased survival (p < 0.01); however, in multivariate analysis, discordant biomarker expression was not associated with decreased survival (p > 0.05). Tumor expression of ER/PR/HER2 can differ between the PT and MET. Loss of ER/PR expression is common and may be related to worse survival. Routine assessment of MET tumor markers could inform prognosis and therapeutic decision-making.

Organoids as a new approach for improving pediatric cancer research.

A key challenge in cancer research is the meticulous development of models that faithfully emulates the intricacies of the patient scenario, with emphasis on preserving intra-tumoral heterogeneity and the dynamic milieu of the tumor microenvironment (TME). Organoids emerge as promising tool in new drug development, drug screening and precision medicine. Despite advances in the diagnoses and treatment of pediatric cancers, certain tumor subtypes persist in yielding unfavorable prognoses. Moreover, the prognosis for a significant portion of children experiencing disease relapse is dismal. To improve pediatric outcome many groups are focusing on the development of precision medicine approach. In this review, we summarize the current knowledge about using organoid system as model in preclinical and clinical solid-pediatric cancer. Since organoids retain the pivotal characteristics of primary parent tumors, they exert great potential in discovering novel tumor biomarkers, exploring drug-resistance mechanism and predicting tumor responses to chemotherapy, targeted therapy and immunotherapies. We also examine both the potential opportunities and existing challenges inherent organoids, hoping to point out the direction for future organoid development.

Tumor biomarkers for diagnosis, prognosis and targeted therapy.

Tumor biomarkers, the substances which are produced by tumors or the body's responses to tumors during tumorigenesis and progression, have been demonstrated to possess critical and encouraging value in screening and early diagnosis, prognosis prediction, recurrence detection, and therapeutic efficacy monitoring of cancers. Over the past decades, continuous progress has been made in exploring and discovering novel, sensitive, specific, and accurate tumor biomarkers, which has significantly promoted personalized medicine and improved the outcomes of cancer patients, especially advances in molecular biology technologies developed for the detection of tumor biomarkers. Herein, we summarize the discovery and development of tumor biomarkers, including the history of tumor biomarkers, the conventional and innovative technologies used for biomarker discovery and detection, the classification of tumor biomarkers based on tissue origins, and the application of tumor biomarkers in clinical cancer management. In particular, we highlight the recent advancements in biomarker-based anticancer-targeted therapies which are emerging as breakthroughs and promising cancer therapeutic strategies. We also discuss limitations and challenges that need to be addressed and provide insights and perspectives to turn challenges into opportunities in this field. Collectively, the discovery and application of multiple tumor biomarkers emphasized in this review may provide guidance on improved precision medicine, broaden horizons in future research directions, and expedite the clinical classification of cancer patients according to their molecular biomarkers rather than organs of origin.

The HER2 flip-HER2 amplification of tumor cells in the cerebrospinal fluid of breast cancer patients with leptomeningeal disease: implications for treating the LM tumor with anti-HER2 therapy.

CNSide is a platform that detects and characterizes tumor cells in the cerebrospinal fluid (CSF) of patients with leptomeningeal disease (LMD). The platform was validated per College of American Pathologists (CAP) and Clinical Laboratories Improvement Amendment (CLIA) guidelines and run as a commercial Laboratory Developed Test (LDT) at Biocept in San Diego, CA. The platform allows CSF tumor cell (CSF-TC) enumeration and biomarker characterization by fluorescent in situ hybridization (FISH). We performed a multicenter retrospective chart review of HER2 FISH CNSide test results that were commercially ordered on 26 patients by physicians for LMD breast cancer patients between April 2020 and October 2022. We show that HER2 is amplified on CSF tumor cells in 62% (16/26) of LMD breast cancer patients. 10/26 (38%) patients had discordant HER2-positivity between the primary tumor tissue and CSF-TC; of these, 35% (9/26) of the patients displayed HER2 amplification on the CSF-TCs, however were categorized as HER2 negative on the primary tumor. Of the 27% (7/26) patients with a HER2 positive primary tumor, one patient showed a HER2 negative LMD tumor. Two patients, 8% (2/26) had a HER2 equivocal primary tumor; of these, one demonstrated a HER2 negative, and one a HER2 positive LMD tumor. Serial analysis (at least 4 longitudinal tests) of HER2 status of the CSF-TC throughout therapy was available for 14 patients and demonstrated that HER2 status of the LMD changed in 29% (4/14) during their treatment course and impacted care decisions. Our data suggests that CSF-TC HER2 FISH analysis in LMD breast cancer patients may be discordant to the primary tumor sample and the discovery of HER2 positivity in the CSF may open doors to anti-HER2 targeted therapy options for LMD patients.