Serum Tumor Biomarkers Research Articles

Baseline computed tomography imaging findings could assist in early diagnosis of visceral pleural invasion for newly discovered early subpleural non-small cell lung cancer: T1 or T2.

Preoperative accurate visceral pleural infiltration (VPI) diagnosis for T1-size non-small cell lung cancer (NSCLC) is significant for clinical decision-making. The study aimed to explore the diagnostic efficacy of computed tomography (CT) imaging features and serum biomarkers in diagnosing VPI in newly discovered subpleural NSCLC ≤3 cm. There were 447 patients with NSCLC ≤3 cm retrospectively enrolled and assigned to the VPI group (n=81) and the non-VPI group (n=366) based on elastic fiber staining results. The serum biomarkers and CT imaging features were obtained for each subject. Univariate and multivariate analyses were used to identify the independent predictors for VPI. Area under the receiver operating characteristic (ROC) curve (AUC) was used to evaluate the diagnostic performance of each independent predictor and combined predictors in predicting VPI, with performance compared using the DeLong test. For tumor biomarkers, the VPI group had a significantly higher percentage of cases with abnormal carcino-embryonic antigen (CEA) level, cytokeratin 19 fragment (CYFRA21-1) level, and pro-gastrin-releasing peptide (ProGRP) level than that of the non-VPI group (P<0.001, P=0.003, P=0.004). However, in multivariate analysis, only the lesion-pleura relationship patterns type Ia [odds ratio (OR) =20.689; 95% confidence interval (CI): 5.058-84.622; P<0.001], type Ib (OR =5.155; 95% CI: 1.178-22.552; P=0.03), type II (OR =7.154; 95% CI: 1.733-29.53; P=0.007) with type III as reference, solid lesion density (OR =9.954; 95% CI: 4.976-19.911; P<0.001) with part-solid density as reference were identified as the independent predictors for VPI. In predicting VPI, the combined model (AUC =0.885) significantly outperformed models based on lesion density (AUC =0.833) and lesion-pleura relationship patterns (AUC =0.655) (all P<0.001). The CT predictors for VPI in patients with subpleural NSCLC (≤3 cm) were lesion density and lesion-pleura relationship patterns (pleural attachment and indentation), but not serum tumor biomarkers.

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.

Abstract CT107: Durable immunogenicity of ELI-002 2P in AMPLIFY-201: Lymph node targeted mKRAS-specific amphiphile vaccine in pancreatic and colorectal cancer

Abstract Background: Colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC) are the second and third leading causes of cancer death in the United States. Kirsten rat sarcoma driver mutations (mKRAS) are prevalent in CRC (52%) and PDAC (93%), making them attractive targets for immunotherapy where therapeutic options are otherwise limited. ELI-002 2P immunotherapy enhances lymph node delivery and immune responses using Amphiphile (Amph) modification of vaccine components. 84% of patients (21/25) induced mKRAS-specific T-cells post-vaccination as assessed by direct ex vivo Fluorospot and/or ICS assays with an average 58-fold increase from baseline. A balanced CD4+ and CD8+ T cell response was observed in 59% of patients. Reductions in ctDNA or serum tumor biomarker antigen from baseline were observed in 84% of patients (21/25) and 24% (6/25) had complete biomarker clearance. The induction of strong mKRAS T cell responses correlated with reductions in tumor biomarker response/clearance and relapse-free survival (RFS) in patients. Methods: ELI-002-001 is a first-in-human Phase 1 trial (NCT04853017) of ELI-002 2P cancer vaccine as adjuvant treatment for patients with high relapse-risk mKRAS+ PDAC and CRC. ELI-002 2P consists of 2 Amph-modified mKRAS peptide antigens, Amph-G12D and Amph-G12R (Amph-Peptides 2P), and an Amph-modified immune-stimulatory oligonucleotide adjuvant (Amph-CpG-7909). 25 patients received ELI-002 2P at 1.4 mg of Amph-Peptides 2P and Amph-CpG-7909 at 5 dose levels; 0.1, 0.5, 2.5, 5, and 10 mg. Peripheral blood and circulating tumor DNA (ctDNA) or serum tumor antigen were collected longitudinally to assess T cell immunogenicity and reductions in clinical biomarkers. Results: A majority of patients who received the booster immunizations maintained or increased mKRAS-specific T cell responses relative to baseline. A high frequency of polyfunctional cells secreting IFNγ, TNFα, IL-2, and/or Granzyme B was observed. CD4+ T regulatory cells were not induced after ELI-002 2P immunization. Assessment of the breadth of responses to 7 different KRAS mutations revealed broad cross-reactivity to KRAS mutants, including non-immunizing epitopes, and no responses to WT. Additional phenotypic and functional qualities were assessed. Conclusions: ELI-002 is an off-the-shelf vaccine targeting common KRAS tumor mutations demonstrating several key advantages: lymph node-targeted vaccine design, high immunogenicity, with balanced CD4+ and CD8+ T cell responses, HLA-agnostic activity, and targeting of vaccine antigens critical for tumor survival. A randomized Phase 2 clinical trial (NCT05726864) investigating a 7-peptide formulation (G12D, R, V, A, C, S, G13D) is underway. Citation Format: James R. Perry, Haley VanWyk, Amy M. Tavares, Thian Kheoh, Esther Welkowsky, Christopher M. Haqq, Peter C. DeMuth, Lisa K. McNeil. Durable immunogenicity of ELI-002 2P in AMPLIFY-201: Lymph node targeted mKRAS-specific amphiphile vaccine in pancreatic and colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr CT107.

Extracellular vesicles-derived CXCL4 is a candidate serum tumor biomarker for colorectal cancer

Extracellular vesicles (EVs) were promising circulating biomarkers for multiple diseases, but whether serum EVs-derived proteins could be used as a reliable tumor biomarker for colorectal cancer (CRC) remained inconclusive. In this study, we identified CXCL4 by a 4D data-independent acquisition-based quantitative proteomics assay of serum EVs-derived proteins in 40 individuals and subsequently analyzed serum EVs-derived CXCL4 levels by ELISA in 2 cohorts of 749 individuals. The results revealed that EVs-derived CXCL4 levels were dramatically elevated in CRC patients than in benign colorectal polyp patients or healthy controls (HC). Furthermore, receiver operating characteristic curves revealed that EVs-derived CXCL4 exhibited superior diagnostic performance with area under the curve of 0.948 in the training cohort. Additionally, CXCL4 could effectively distinguish CRC in stage I/II from HC. Notably, CRC patients with high levels of EVs-derived CXCL4 have shorter 2-year progression-free survival than those with low levels. Overall, our findings demonstrated that serum EVs-derived CXCL4 was a candidate diagnostic and prognostic biomarker for CRC.

Comparing modeling strategies combining changes in multiple serum tumor biomarkers for early prediction of immunotherapy non-response in non-small cell lung cancer.

Patients treated with immune checkpoint inhibitors (ICI) are at risk of adverse events (AEs) even though not all patients will benefit. Serum tumor markers (STMs) are known to reflect tumor activity and might therefore be useful to predict response, guide treatment decisions and thereby prevent AEs. This study aims to compare a range of prediction methods to predict non-response using multiple sequentially measured STMs. Nine prediction models were compared to predict treatment non-response at 6-months (n = 412) using bi-weekly CYFRA, CEA, CA-125, NSE, and SCC measurements determined in the first 6-weeks of therapy. All methods were applied to six different biomarker combinations including two to five STMs. Model performance was assessed based on sensitivity, while model training aimed at 95% specificity to ensure a low false-positive rate. In the validation cohort, boosting provided the highest sensitivity at a fixed specificity across most STM combinations (12.9% -59.4%). Boosting applied to CYFRA and CEA achieved the highest sensitivity on the validation data while maintaining a specificity >95%. Non-response in NSCLC patients treated with ICIs can be predicted with a specificity >95% by combining multiple sequentially measured STMs in a prediction model. Clinical use is subject to further external validation.

Combined use of CYFRA 21-1 and CA 125 predicts survival of patients with metastatic NSCLC and stable disease in IMpower150.

Patients with non-small cell lung cancer (NSCLC) and stable disease (SD) have an unmet clinical need to help guide early treatment adjustments. To evaluate the potential of tumor biomarkers to inform on survival outcomes in NSCLC SD patients. This post hoc analysis included 480 patients from the IMpower150 study with metastatic NSCLC, treated with chemotherapy, atezolizumab and bevacizumab combinations, who had SD at first CT scan (post-treatment initiation). Patients were stratified into high- and low-risk groups (overall survival [OS] and progression-free survival [PFS] outcomes) based on serum tumor biomarker levels. The CYFRA 21-1 and CA 125 biomarker combination predicted OS and PFS in patients with SD. Risk of death was ~4-fold higher for the biomarker-stratified high-risk versus low-risk SD patients (hazard ratio [HR] 3.80; 95% confidence interval [CI] 3.02-4.78; p < 0.0001). OS in patients with the low- and high-risk SD was comparable to that in patients with the CT-defined partial response (PR; HR 1.10; 95% CI 0.898-1.34) and progressive disease (PD) (HR 1.05; 95% CI 0.621-1.77), respectively. The findings were similar with PFS, and consistent across treatment arms. Biomarker testing shows potential for providing prognostic information to help direct treatment in NSCLC patients with SD. Prospective clinical studies are warranted.ClinicalTrials.gov: NCT02366143.

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

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. 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. 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 < 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 < 0.05). 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.

Molecular Biomarkers in Cholangiocarcinoma: Focus on Bile.

Hepatobiliary system cancers have demonstrated an increasing incidence rate in the past years. Without the presence of early symptoms, the majority of such cancers manifest with a set of similar symptoms, such as cholestasis resulting in posthepatic icterus. Differential diagnosis of hepatobiliary cancers is required for the therapy selection, however, the similarity of the symptoms complicates diagnostics. Thus, the search for molecular markers is of high interest for such patients. Cholangiocarcinoma (CCA) is characterized by a poor prognosis due to a low resectability rate, which occurs because this disease is frequently beyond the limits of surgical therapy at the time of diagnosis. The CCA is diagnosed by the combination of clinical/biochemical features, radiological methods, and non-specific serum tumor biomarkers, although invasive examination is still needed. The main disadvantage is limited specificity and sensitivity, which complicates early diagnostics. Therefore, prognostic and predictive biomarkers are still lacking and urgently needed for early diagnosis. In contrast to serum, bile is more accessible to identify biliary disease due to its simpler composition. Moreover, bile can contain higher concentrations of tumor biomarkers due to its direct contact with the tumor. It is known that the composition of the main bile component - bile acids, may vary during different diseases of the biliary tract. This review summarizes the recent developments in the current research on the diagnostic biomarkers for CCA in serum and bile and provides an overview of the methods of bile acids analysis.

A combination of radiomic features, clinic characteristics, and serum tumor biomarkers to predict the possibility of the micropapillary/solid component of lung adenocarcinoma.

Invasive lung adenocarcinoma with MPP/SOL components has a poor prognosis and often shows a tendency to recurrence and metastasis. This poor prognosis may require adjustment of treatment strategies. Preoperative identification is essential for decision-making for subsequent treatment. This study aimed to preoperatively predict the probability of MPP/SOL components in lung adenocarcinomas by a comprehensive model that includes radiomics features, clinical characteristics, and serum tumor biomarkers. A retrospective case control, diagnostic accuracy study. This study retrospectively recruited 273 patients (males: females, 130: 143; mean age ± standard deviation, 63.29 ± 10.03 years; range 21-83 years) who underwent resection of invasive lung adenocarcinoma. Sixty-one patients (22.3%) were diagnosed with lung adenocarcinoma with MPP/SOL components. Radiomic features were extracted from CT before surgery. Clinical, radiomic, and combined models were developed using the logistic regression algorithm. The clinical and radiomic signatures were integrated into a nomogram. The diagnostic performance of the models was evaluated using the area under the curve (AUC). Studies were scored according to the Radiomics Quality Score and Transparent Reporting of a Multivariable Prediction Model for Individual Prognosis or Diagnosis guidelines. The radiomics model achieved the best AUC values of 0.858 and 0.822 in the training and test cohort, respectively. Tumor size (T_size), solid tumor size (ST_size), consolidation-to-tumor ratio (CTR), years of smoking, CYFRA 21-1, and squamous cell carcinoma antigen were used to construct the clinical model. The clinical model achieved AUC values of 0.741 and 0.705 in the training and test cohort, respectively. The nomogram showed higher AUCs of 0.894 and 0.843 in the training and test cohort, respectively. This study has developed and validated a combined nomogram, a visual tool that integrates CT radiomics features with clinical indicators and serum tumor biomarkers. This innovative model facilitates the differentiation of micropapillary or solid components within lung adenocarcinoma and achieves a higher AUC, indicating superior predictive accuracy.

Abstract C092: T cell responses and clinical outcomes in pancreatic and colorectal cancer patients with minimal residual disease in AMPLIFY-201, a phase 1 trial of a first-in-class amphiphile lymph node targeted mutant KRAS vaccine

Abstract Background: Mutations in RAS occur in &amp;gt; 25% of solid tumors and &amp;gt; 90% of patients (pts) with PDAC and &amp;gt; 40% of pts with CRC. ELI-002 2P is a vaccine comprised of lymph-node targeted Amphiphile (Amph)-modified G12D and G12R mutant KRAS peptides together with an Amph-modified CpG oligonucleotide adjuvant designed to expand polyfunctional mutant KRAS-specific T cells. Interim phase 1 AMPLIFY-201 data (O’Reilly et al; ASCO, 2023, #2528) showed that adjuvant administration of ELI-002 was well-tolerated, with T cell responses in 87%, and tumor biomarker responses in 77% (a subset of 32% had ctDNA clearance). Here we report relapse free survival data and correlations between ELI-002 2P- induced T cell levels and outcomes in pts with PDAC and colorectal cancer (CRC) at high risk for relapse following locoregional treatment. Methods: Resected PDAC (n=20) and CRC (n=5) pts with tumors harboring KRAS G12D or G12R who had minimal residual disease positivity (MRD+) defined as elevated circulating tumor DNA (ctDNA) and/or serum tumor biomarker (CA19-9/CEA), received up to 6 priming doses and 4 booster doses separated by a 3-month rest period of subcutaneous ELI-002 2P vaccine monotherapy comprised of Amph-peptides (700 mcg each G12D/G12R), admixed with Amph-CpG-7909 at 0.1, 0.5, 2.5, 5.0 and 10.0 mg per cohort dose level. Primary endpoints included safety and recommended phase 2 dose (RP2D) of Amph-CPG-7909; secondary endpoints: biomarker reduction/clearance; exploratory endpoints: included relapse free survival (RFS) using immune Response Evaluation Criteria in Solid Tumors (iRECIST) and immunogenicity assessed by direct ex vivo Fluorospot and intracellular cytokine staining of peripheral blood mononuclear cells. Results: Direct ex vivo polyfunctional mKRAS-specific T cell responses to ELI-002 2P were observed in 20/23 (87%; 50% induced both CD4+ and CD8+ T cells, median 13-fold and mean 56-fold increase from baseline), with response in 9/9 (100%) pts treated at highest two dose levels including the 10 mg RP2D. Clinical efficacy correlated with T cell response in evaluable pts (n=22): median tumor biomarker reduction/clearance was -86.9% vs -4.5% in above vs below median T cell responders, respectively (p &amp;lt; 0.0124). At 25 weeks median follow-up, the median RFS was not reached compared to 17.1 weeks in above versus below median T cell responders (HR 0.11; 95% CI 0.027-0.446; p = 0.0108). The association of RFS with T cell response was not confounded by other baseline prognostic variables (including tumor stage, recovery from prior cytotoxic therapy as assessed by absolute neutrophil count, or immune system subsets such as %CD4+ or %CD8+of CD3+ lymphocytes). Conclusions: The association of ELI-002 2P vaccine T cell responses with clinical outcomes in a novel MRD+ trial design underscores the potential to reduce recurrence in the setting of microsatellite stable PDAC and CRC. A new phase 1/2 trial, AMPLIFY-7P (NCT05726864), is evaluating a new seven peptide (G12D, G12R, G12V, G12C, G12A, G12S, G13D) ELI-002 7P formulation. Citation Format: Zev A. Wainberg, Shubham Pant, Colin D. Weekes, Muhammad Furqan, Pashtoon M. Kasi, Craig E. Devoe, Alexis D. Leal, Vincent Chung, James R. Perry, Lochana Seenappa, Lisa K. McNeil, Esther Welkowsky, Peter C. DeMuth, Christopher M. Haqq, Eileen M. O'Reilly. T cell responses and clinical outcomes in pancreatic and colorectal cancer patients with minimal residual disease in AMPLIFY-201, a phase 1 trial of a first-in-class amphiphile lymph node targeted mutant KRAS vaccine [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr C092.

Abstract B039: A phase 1/2 study of dendrimer-enhanced (DEP) SN38 (SN38-SPL9111/DEP irinotecan) in patients with advanced solid tumours

Abstract Background: Dendrimers are highly branched nanoparticles that achieve significant tumor targeting of cytotoxic drugs. DEP SN38 (or DEP irinotecan) is a highly optimised dendrimer conjugate of the irinotecan active metabolite, SN38. We report initial results of a phase 1/2 trial evaluating the safety, tolerability and preliminary efficacy of DEP SN38 in patients (pts) with advanced solid tumors, including colorectal (CRC) and platinum-resistant high-grade serous ovarian carcinoma (HGSOC). Methods: Solid tumor pts who had exhausted standard therapy were enrolled (EudraCT2019-001318-40). DEP SN38 given IV 3-weekly (Q3W) was escalated to identify a recommended phase 2 dose (RP2D). Additional dose assessment/expansion cohorts followed at Q3W or Q2W (2-weekly) alone or in combination with 5-fluorouracil and leucovorin (5FU/LV). Efficacy was assessed via RECIST 1.1 criteria and serum tumor biomarkers. Results: Dose escalation (N=7) confirmed a Q3W RP2D of 12.5 mg/m2 SN38 with 1 pt having a dose limiting toxicity (DLT) of grade 4 neutropenia for &amp;gt;7d. Following successful dose expansion, additional dose exploration confirmed a Q2W RP2D of 12.5 mg/m2 SN38 alone or in combination with 5FU/LV. DEP SN38 was well-tolerated for all dose regimens (N=101) with mostly mild/moderate treatment-related adverse events (TRAEs), with no new events compared to those observed with conventional irinotecan. Notably, there were no reports of severe (≥ grade 3) diarrhea with DEP SN38 and no reports of cholinergic symptoms, both being common with conventional irinotecan (~20% and 47% pts, respectively). Moreover, with DEP SN38, severe nausea (2.2% pts) and vomiting (1.1% pts) occurred less frequently than with irinotecan (both ~10% pts). While febrile neutropenia was observed as a DLT at 15 mg/m2 Q2W monotherapy (2 pts), neutropenia was otherwise uneventful and managed with G-CSF.CRC pts: 38 heavily pre-treated CRC pts were dosed (N=31 evaluable); pts had an average ~4 prior treatment lines with 97% having received at least 1 irinotecan containing line. DEP SN38 achieved a disease control rate (DCR) of 48%, with stable disease up to 72 weeks.HGSOC pts: 23 platinum-resistant HGSOC pts were dosed (N=18 evaluable); pts had received an average ~6 prior treatment lines. DCR for pts dosed Q2W was 100% with 33.3% objective response rate, and 72% DCR for all HGSOC pts. There were 3 partial responses (PR), for up to 27 wks with 1 pt achieving complete tumour and ascites resolution with pts ongoing. 75% of pts achieved CA-125 reductions of up to 98% vs baseline. For DEP SN38 in combination with 5FU/LV, in a cohort of mainly CRC pts (N=6), DCR is 100% including 1 PR, with pts ongoing. Conclusions: DEP SN38 was very well-tolerated with significantly fewer severe gastrointestinal TRAEs compared to conventional irinotecan. Encouraging anti-tumour activity, including prolonged disease control in heavily pre-treated CRC and HGSOC pts, demonstrates promising clinical utility of DEP SN38, both as a monotherapy and as a combination therapy. Recruitment is ongoing. Citation Format: Jia Liu, Anna R Minchom, Alastair Greystoke, Thomas R J Evans, Debashis Sarker, Anthony M Joshua, Cienne Morton, Aisha Gaus, Wing Yau, Rasha Cosman, Dominika Chwialkowska, Jeremy R A Paull, Bernadette M Jean-Francois, Jacinth K Fairley, Nicola J Main, Stephanie R Edmondson, Natalie Cook. A phase 1/2 study of dendrimer-enhanced (DEP) SN38 (SN38-SPL9111/DEP irinotecan) in patients with advanced solid tumours [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr B039.

Serial circulating tumor DNA profiling predicts tumor recurrence after liver transplantation for liver cancer.

Minimal residual disease (MRD) is proposed to be responsible for tumor recurrence. The role of circulating tumor DNA (ctDNA) to detect MRD, monitor recurrence, and predict prognosis in liver cancer patients undergoing liver transplantation (LT) remains unrevealed. Serial blood samples were collected to profile ctDNA mutational changes. Baseline ctDNA mutational profiles were compared with those of matched tumor tissues. Correlations between ctDNA status and recurrence rate (RR) and recurrence-free survival (RFS) were analyzed, respectively. Dynamic change of ctDNA was monitored to predict tumor recurrence. Baseline mutational profiles of ctDNA were highly concordant with those of tumor tissues (median, 89.85%; range 46.2-100%) in the 74 patients. Before LT, positive ctDNA status was associated with higher RR (31.7% vs 11.5%; p = 0.001) and shorter RFS than negative ctDNA status (17.8 vs 19.4months; p = 0.019). After LT, the percentage of ctDNA positivity decreased (17.6% vs 47.0%; p < 0.001) and patients with positive ctDNA status had higher RR (46.2% vs 21.3%; p < 0.001) and shorter RFS (17.2 vs 19.2months; p = 0.010). Serial ctDNA profiling demonstrated patients with decreased or constant negative ctDNA status had lower RR (33.3% vs 50.0%; p = 0.015) and favorable RFS (18.2 vs 15.0months, p = 0.003) than those with increased or constant positive ctDNA status. Serial ctDNA profiling predicted recurrence months ahead of imaging evidence and serum tumor biomarkers. ctDNA could effectively detect MRD and predict tumor recurrence in liver cancer patients undergone LT.

Serum tumor biomarkers as a surrogate for radiographic assessment of non-small cell lung cancer.

e21076 Background: Conventional tumor markers are positioned to serve as a useful adjunct in lung cancer management. However, most studies in this field have been specific to a particular disease stage or treatment regimen. Thus, the purpose of this research was to assess whether three minimally invasive, low-cost serum tumor markers (CEA, CA19-9, and CA-125) held associations with radiographic and clinical outcomes in non-small cell lung cancer (NSCLC) patients who received systemic therapy in a more comprehensive patient population. Methods: This was a single-center retrospective study of NSCLC patients treated between January 2016 and August 2020. Serum tumor markers were statistically analyzed for differences in patients who responded or progressed (RECIST 1.1 or iRECIST criteria), associations with demographic and clinical characteristics, and all-cause mortality in pre-defined populations. Disease response was assessed radiographically using RECIST 1.1 criteria. Results: From 533 NSCLC patients screened, 165 met inclusion criteria. Of these, 50.9% were male and 49.1% female. 69.7% had stage IV disease at baseline. The proportion of patients with an elevated CEA, CA-125, and CA19-9 at baseline were 58.8%, 50.9%, and 30.3%, respectively. A subset of 92 patients had paired tumor markers and radiographic scans, from which median (IQR) fold-change in tumor markers from nadir to progression was 2.13 (IQR 1.24 - 3.02; p &lt; 0.001) for CEA (n = 47), 1.46 (IQR 1.13 - 2.18; p &lt; 0.001) for CA19-9 (n = 46), and 1.53 (IQR 0.96 - 2.12; p &lt; 0.001) for CA-125 (n = 47). Median (IQR) fold-change in tumor markers from baseline to radiographic response was 0.50 (IQR 0.27, 0.95; p &lt; 0.001) for CEA (n = 39), 1.08 (IQR 0.74, 1.61; p = 0.99) for CA19-9 (n = 35), and 0.47 (IQR 0.18, 1.26; p = 0.008) for CA-125 (n = 35). Lastly, an elevated baseline CEA was not associated with a difference in overall survival (HR 1.05, 95% CI 0.63-1.74, p = 0.84) in patients with stage IV disease within the total population. Conclusions: Serum CEA, CA-125, and CA19-9 levels were significantly different than nadir in patients who progressed. CEA and CA-125 were significantly different than baseline in those who responded. These cost-effective tumor markers may serve as an important adjunct to clinical decision making. Analysis within a controlled clinical trial is warranted.

AMPLIFY-201, a first-in-human safety and efficacy trial of adjuvant ELI-002 2P immunotherapy for patients with high-relapse risk with KRAS G12D- or G12R-mutated pancreatic and colorectal cancer.

2528 Background: RAS mutations occur in 25% of solid tumors with G12D being the most frequent variant. ELI-002 2P is a vaccine comprised of lymph-node targeted Amphiphile (Amph)-modified G12D and G12R mutant KRAS peptides with an Amph-modified CpG oligonucleotide adjuvant designed to expand polyfunctional mutant KRAS-specific T cells. Preclinical amphiphile data (relative to non-amphiphile controls) showed increased immunogenicity, tumor clearance and survival in mouse models. Methods: This first-in-human multicenter phase I trial assessed safety, immunogenicity and antitumor activity using a novel adjuvant trial design in patients (pts) with minimal residual disease (MRD) following standard locoregional treatment. Eligibility: elevated circulating tumor DNA (ctDNA) and/or serum tumor biomarker elevation (CA19-9/CEA), and KRAS/NRAS mutation. In a 3+3 dose escalation, n=22 received a subcutaneous course of 6 prime and 4 booster injections of fixed dose Amph-peptides (1.4 mg), admixed with escalating Amph-CpG-7909 (Table). Dose escalation based on observed safety in first 28 days. Safety, antitumor activity including biomarker reduction/clearance and relapse free survival using immune Response Evaluation Criteria in Solid Tumors (iRECIST) were assessed. Immunogenicity assessed by direct ex vivo Fluorospot and intracellular cytokine staining of peripheral blood mononuclear cells (T-cell responders = ≥ 2-fold increase over baseline) and by tumor CD3 immunohistochemistry in subset who underwent a biopsy. Results: No dose-limiting toxicities, treatment related SAEs or cytokine release syndrome were observed, and no maximum tolerated dose was identified. Safety: all grade 1, fatigue (19%), headache (19%), and injection site reaction (9.5%). Biomarker reduction was observed in 15/19 (79%) and in 5/5 at highest doses; clearance of MRD was observed in 4/19 (21% - n=2 pancreas, n=2 colorectal). Polyfunctional mKRAS-specific T cell responses observed in 80% of pts (n=12/15), with both CD8+ and CD4+ T cell responses for most, and CD3+ T cells observed on biopsies. The recommended phase 2 dose (RP2D) is 10.0 mg Amph-CpG-7909. Conclusions: ELI-002 2P, evaluated in a novel MRD+ trial, was safe with ctDNA and serum tumor biomarker reduction and clearance and notable immune responses. The RP2D is the start dose for a phase 1/randomized phase 2 study evaluating a new seven peptide formulation for G12 D, R, V, S, A, C, and G13D mutated solid tumors (NCT05726864). Clinical trial information: NCT04853017 . [Table: see text]

HSD32 Are We ‘Choosing Wisely’? Use of Surveillance Advanced Imaging and Serum Tumor Biomarker Testing in Older Women with Breast Cancer

The American Society of Clinical Oncology’s (ASCO) Choosing Wisely (CW) initiative recommends against the use of surveillance testing (ST) of asymptomatic women with breast cancer (BC) using advanced imaging or serum tumor biomarkers testing. We evaluated if CW impacted the use of ST in women with BC. Also, its predictors were identified.

Role of CA 19.9 and CEA in predicting diagnosis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is responsible for 90% of cases of primary liver cancer and is also responsible for the fourth most common cause of cancer death worldwide. To improve the current scenario for the early diagnosis and management of HCC patients, a better understanding of HCC is required. Hence, serum tumor biomarkers such as CA 19-9 (cancer antigen), CEA (carcinoembryonic antigen), and AFP (alpha-fetoprotein) show promising future, when it comes to early diagnosis of primary liver cancer (PHC), liver cirrhosis, and metastasis. It was a retrospective cross-sectional analysis of patients diagnosed with primary hepatocellular carcinoma, data were collected from the hospital database and included a total of 245 patients of HCC attending the out-patient department and some were admitted for treatment at our institution. Out of 245 patients, 68 patients were selected for the study. We have collected information related to the patient's demographic profile, pathological diagnosis, biochemical profile, and even radiological diagnosis. The sensitivity and specificity of CA 19-9 and CEA were also done. Adenocarcinoma is the most common type of liver cancer followed by HCC. We have shown a weak correlation between tumor markers CA 19-9 and CEA for the diagnosis of liver carcinoma. Further our study shows that the sensitivity of tumor marker CA 19-9 for the diagnosis of liver carcinoma is 64.28% and that of CEA is 83.67%. The search for a novel biomarker of early liver carcinoma requires further research. The authors declare that they have no competing interests.

The value of folate receptor-positive circulating tumor cells in the diagnosis of lung cancer and its correlation with clinical characteristics.

The aim of this research is to investigate the feasibility of folate receptor-positive circulating tumor cells (FR+CTCs) as a biomarker for the diagnosis of malignant pulmonary nodules and the correlation between clinicopathological factors and FR+CTC levels. Patients initially diagnosed with one or more pulmonary nodules from a computed tomography scan were prospectively included. Three milliliters of peripheral blood was collected from each participant for FR+CTC analysis prior to surgery. Clinical and pathological parameters and FR+CTC levels were compared between patients with lung cancer and benign diseases. Six hundred fifty-three patients had lung cancer and the other 124 had benign lung diseases based on pathological examinations of the resected specimens. The median FR+CTC value of the lung cancer group was 12.0(95% CI 9.6-16.2)FU/3mL and that of the benign group was 7.2(95% CI 5.78-11.2)FU/3mL. The difference was statistically significant (P < 0.0001). In a receiver operating characteristic analysis to distinguish the two groups, the area under curve of FR+CTC was 0.7457 (95% CI 0.6893-0.8021; P < 0.0001) using a cutoff of 8.65 FU/3mL. The sensitivity was 86.37%, and the specificity was 74.19%. Combined with conventional serum tumor biomarkers, the area under curve was 0.922 (0.499-0.963). The sensitivity was 92.20%, and the specificity was 83.05%. FR+CTC levels were related to tumor staging (P4< 0.001), the degree of tumor invasion both in single (P = 0.011) and multiple lesions (P = 0.022), pathological subtypes (P = 0.013), and maximum tumor diameter (P = 0.014). FR+CTC is an effective and reliable biomarker for the diagnosis of lung cancer. Further, FR+CTC level is correlated with tumor staging, degree of invasion, pathological subtypes, and tumor size.

Diagnostic value of aberrant decreased 5-Methylcytosine RNA modification in leukocytes for non-small cell lung cancer.

Background: Non-small cell lung cancer (NSCLC) was a disease with poor outcomes, partly because there were no high-efficiency non-invasive diagnostic biomarkers. The RNA modification status of 5-Methylcytosine (m5C) has been shown to be a biomarker for various diseases, but its potentiality to be a diagnostic biomarker for NSCLC remained inconclusive. Methods: In this research, we collected peripheral leukocyte samples from 141 patients with NSCLC and 90 normal people as controls to evaluate the extent of m5C RNA modification. Results: We found that the m5C modification levels in leukocytes of NSCLC patients were decreased dramatically, which were compared to the normal controls, and levels of m5C modification decreased progressively with tumor stage. Importantly, m5C modification exhibited superior diagnostic value compared to carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), cytokeratin 19 fragment (Cyfra21-1), and carbohydrate antigen 125 (CA125), which demonstrated area under the curves (AUCs) of 0.912, 0.773, 0.669, 0.754, and 0.732, respectively. The combination of m5C modification with these serum tumor biomarkers further improved the AUC to 0.960. A nomogram model incorporating m5C modification also provided an effectively diagnostic tool for NSCLC. Conclusion: Collectively, our findings suggested that m5C modification in leukocytes held promise as a prospective biomarker for NSCLC diagnosis.

Enrichment of Circulating Tumor Cells of Lung Cancer and Correlation With Serum Leukomonocyte and Tumor Biomarkers: A Retrospective Study.

Circulating tumor cells is important in the clinical diagnosis of cancer and there are a number of circulating tumor cell detection systems associated with different isolation strategies being validated. There is a novel platform, the CytoBot 2000, which utilizes a combination of physical and immunological technologies to isolate and capture circulating tumor cells. In this retrospective study, 39 lung cancer patients and 11 normal healthy individuals were enrolled and performed circulating tumor cell tests and immunofluorescence staining with CytoBot 2000. The performance of this device was assessed by receiver operating characteristic curve. The clinical relevance of circulating tumor cells was assessed by Chi-square. The correlations between circulating tumor cell number and blood lymphocytes and tumor biomarkers were analyzed by Pearson correlation coefficient. The number of circulating tumor cell is significantly increased in lung cancer patients (3.74 > 0.45, P < .0001). The CytoBot 2000 presented a 100% (39/39) circulating tumor cell detection rate in lung cancer patients and 36% (4/11) in healthy individual blood samples, the sensitivity and specificity were 89.7% and 90.9%, respectively, and with the area under curve of 0.966. Further, there was a positive correlation between circulating tumor cell count and carcinoembryonic antigen 211 (R2 = 0.125, P = .027), but not blood lymphocytes (P = .089). This automatic platform showed excellent performance of circulating tumor cell detection by clinical sample. The tumor biomarkers increased with the number of circulating tumor cell in the lung cancer patients.

Detection of Novel Autoantibodies to Nucleolin's RNA-binding Domains as a Serum Tumor Biomarker Through ELISA.

Expression and location of nucleolin are often abnormal in malignancies, which may result in the production of autoantibodies. Despite this, the identification of such autoantibodies may be essential for the early diagnosis and prognosis of cancers. In this investigation, the recombinant nucleolin protein was generated using an Escherichia coli expression system and was used an indirect enzyme-linked immunosorbent assay to detect anti-nucleolin autoantibodies in cancer patients' sera. Lung cancer patients' autoantibodies displayed the highest seroreactivity with the recombinant protein, with area under the curve of 0.948 and sensitivity and specificity of 85% and 96.67%, respectively (accuracy=92%). Anti-nucleolin autoantibodies were linked with lung tumor size (r=0.793), tumor, node, metastasis staging (r=0.643), and proliferation (r=0.744). These autoantibodies distinguished patients with early-stage lung cancer from healthy controls. Since anti-nucleolin autoantibodies are strongly linked to tumor size, clinical staging, and growth, they can be used to measure how well a treatment is working.