Circulating Tumor Cells Test Research Articles

Detection and surveillance of circulating tumor cells in sarcoma for predicting therapy response and prognosis.

e23573 Background: Generally sarcomas are categorized as soft tissue and bone sarcomas and represent heterogenous group of malignancies with more than 70 distinct histologic subtypes. Due to their rarity and heterogeneity, sarcomas present particular challenges for accurate diagnosis, prognosis, and treatment. The presence and persistence of circulating tumor cells (CTCs) in the peripheral blood are believed to be associated with poor prognosis and distant metastases. A blood-based CTCs test is thus greatly needed for monitoring disease progression and predicting clinical outcomes. Methods: In this study, we developed a CTCs test for the detection and surveillance of sarcoma patients’ CTCs, and subsequently explored its clinical value. 123 patients with sarcomas were enrolled as the cohort for serial CTCs tests. Dynamic CTCs counting, in combination with therapy evaluation and post-treatment follow-up, was used to explore predicting pre- and post-chemotherapy evaluation and prognosis. Results: Quantification of CTCs demonstrated a significant increase in sarcoma patients compared to healthy individuals. Furthermore, patients in stage III-IV had a higher CTC count than those in stage I-II (p<0.01), suggesting a correlation between CTCs and disease development. Remarkably, there was variation in the quantity of CTCs among different types of sarcomas. The highest number of CTCs was observed in undifferentiated small round cell sarcoma, followed by soft tissue tumor, while bone tumors had the lowest number. In addition, the number of CTCs in patients with positive PD-L1 expression was considerably greater than in those with negative expression (p<0.01). The count of CTCs shown a substantial decrease following neoadjuvant chemotherapy treatment. There was a decline in the number of CTCs both before and after postoperative adjuvant chemotherapy treatment, however, there was no significant statistical discrepancy. The number of CTCs at the baseline was shown to be considerably larger in the group of patients with progressive disease or stable disease (PD+SD) compared to the group with partial response or complete response (PR+CR) (p<0.05). This difference was particularly pronounced for interstitial-type CTCs (p<0.01). Conclusions: The rate of presence of CTCs in the peripheral blood of sarcoma patients is high, and patients with an increased percentage of CTCs after treatment have a poor treatment effect. The dynamic monitoring of changes in CTCs counts after treatment has clinical significance for the timely detection of recurrence or metastasis. CTCs also showed a clinical value in prediction of therapy efficiency or prognosis, and may be clinically valuable for the whole course management of patients.

An indocyanine green-based liquid biopsy test for circulating tumor cells for pediatric liver cancer.

Hepatoblastoma and HCC are the most common malignant hepatocellular tumors seen in children. The aim of this study was to develop a liquid biopsy test for circulating tumor cells (CTCs) for these tumors that would be less invasive and provide real-time information about tumor response to therapy. For this test, we utilized indocyanine green (ICG), a far-red fluorescent dye used clinically to identify malignant liver cells during surgery. We assessed ICG accumulation in cell lines using fluorescence microscopy and flow cytometry. For our CTC test, we developed a panel of liver tumor-specific markers, including ICG, Glypican-3, and DAPI, and tested it with cell lines and noncancer control blood samples. We then used this panel to analyze whole-blood samples for CTC burden with a cohort of 15 patients with hepatoblastoma and HCC and correlated with patient characteristics and outcomes. We showed that ICG accumulation is specific to liver cancer cells, compared to nonmalignant liver cells, non-liver solid tumor cells, and other nonmalignant cells, and can be used to identify liver tumor cells in a mixed population of cells. Experiments with the ICG/Glypican-3/DAPI panel showed that it specifically tagged malignant liver cells. Using patient samples, we found that CTC burden from sequential blood samples from the same patients mirrored the patients' responses to therapy. Our novel ICG-based liquid biopsy test for CTCs can be used to specifically detect and quantify CTCs in the blood of pediatric patients with liver cancer.

Detection of circulating tumor cells by means of machine learning using Smart-Seq2 sequencing

Circulating tumor cells (CTCs) are tumor cells that separate from the solid tumor and enter the bloodstream, which can cause metastasis. Detection and enumeration of CTCs show promising potential as a predictor for prognosis in cancer patients. Furthermore, single-cells sequencing is a technique that provides genetic information from individual cells and allows to classify them precisely and reliably. Sequencing data typically comprises thousands of gene expression reads per cell, which artificial intelligence algorithms can accurately analyze. This work presents machine-learning-based classifiers that differentiate CTCs from peripheral blood mononuclear cells (PBMCs) based on single cell RNA sequencing data. We developed four tree-based models and we trained and tested them on a dataset consisting of Smart-Seq2 sequenced data from primary tumor sections of breast cancer patients and PBMCs and on a public dataset with manually annotated CTC expression profiles from 34 metastatic breast patients, including triple-negative breast cancer. Our best models achieved about 95% balanced accuracy on the CTC test set on per cell basis, correctly detecting 133 out of 138 CTCs and CTC-PBMC clusters. Considering the non-invasive character of the liquid biopsy examination and our accurate results, we can conclude that our work has potential application value.

Detection and identification of circulating tumor cells in parathyroid tumors and correlation analysis with clinicopathological features.

The differential diagnosis of parathyroid carcinoma (PC)/parathyroid adenoma (PA) in parathyroid tumors is critical for their management and prognosis. Circulating tumor cells (CTCs) identification in the peripheral blood of parathyroid tumors remains unknown. In this study, we proposed to investigate the differences of CTCs in PC/PA and the relationship with clinicopathologic features to assess its relevance to PC and value in identifying PC/PA. Peripheral blood was collected from 27 patients with PC and 37 patients with PA treated in our hospital, and the number of chromosome 8 aberrant CTCs was detected by negative magnetic bead sorting fluorescence in situ hybridization (NE-FISH). The differences of CTCs in PC/PA peripheral blood were compared and their diagnostic efficacy was evaluated, and the correlation between CTCs and clinicopathological features of PC was further explored. CTCs differed significantly in PC/PA (p = 0.0008) and were up-regulated in PC, with good diagnostic efficacy. CTCs combined with alkaline phosphatase (ALP) assay improved the diagnostic efficacy in identifying PC/PA (AUC = 0.7838, p = 0.0001). The number of CTCs was correlated with tumor dimensions, but not significantly correlated with clinical markers such as calcium and PTH and pathological features such as vascular invasion, lymph node metastasis and distant metastasis. As a non-invasive liquid biopsy method, CTCs test combined with ALP test can be used as an important reference basis for timely and accurate identification and treatment of PC. It is of great significance to improve the current situation of PC diagnosis, treatment and prognosis.

Circulating tumor cells help differentiate benign ovarian lesions from cancer before surgery: A literature review and proof of concept study using flow cytometry with fluorescence imaging.

Current tools are insufficient for distinguishing patients with ovarian cancer from those with benign ovarian lesions before extensive surgery. The present study utilized a readily accessible platform employing a negative selection strategy, followed by flow cytometry, to enumerate circulating tumor cells (CTCs) in patients with ovarian cancer. These counts were compared with those from patients with benign ovarian lesions. CTC counts at baseline, before and after anticancer therapy, and across various clinical scenarios involving ovarian lesions were assessed. A negative-selection protocol we proposed was applied to patients with suspected ovarian cancer and prospectively utilized in those subsequently confirmed to have malignancy. The protocol was implemented before anticancer therapy and at months 3, 6, 9 and 12 post-treatment. A cut-off value for CTC number at 4.75 cells/ml was established to distinguish ovarian malignancy from benign lesions, with an area under the curve of 0.900 (P<0.001). In patients with ovarian cancer, multivariate Cox regression analysis revealed that baseline CTC counts and the decline in CTCs within the first three months post-therapy were significant predictors of prolonged progression-free survival. Additionally, baseline CTC counts independently prognosticated overall survival. CTC counts obtained with the proposed platform, used in the present study, suggest that pre-operative CTC testing may be able to differentiate between malignant and benign tumors. Moreover, CTC counts may indicate oncologic outcomes in patients with ovarian cancer who have undergone cancer therapies.

Abstract 7493: Real-time monitoring of CTC number, biomarker expression, and kinetics as prognostic and predictive markers in patients with GI malignancies

Abstract Background: Most of the utility in the clinic regarding liquid biopsies has been around circulating tumor DNA (ctDNA). ctDNA analysis provides valuable insight into mutational burden and therapeutic targets, but the speed and cost of analysis are prohibitive especially when it comes to serial testing. Using a cohort of patients with advanced or metastatic gastrointestinal (GI) cancers, we herein show the feasibility of using circulating tumor cell (CTC) enumeration, biomarker analysis, and kinetics as complementary to or as an alternative to ctDNA testing. Methods: Using the RareCyte platform, nucleated cells from blood were collected, spread on slides, stained, and imaged with CyteFinder® for CTC identification (CK/EpCAM+, CD45-) and quantification of biomarkers (PD-L1/HER2 or Ki67/EGFR). CTC enumeration (CLIA-accredited CTC test) and biomarker status for each patient were tracked over time and compared with ctDNA mutational analysis and tumor burden. Results: CTCs were detected in 60/70 samples (86%) and in 36/38 unique patients (95%). On average, more CTCs were detected in baseline samples from colorectal cancer patients compared to other subtypes (Table 1). Moreover, multiple patients showed pronounced changes in CTC number that correlated with treatment response, as well as strong correlation between CTC and tissue biomarker expression e.g. HER2. Rapid decline and/or CTC clearance was seen as early as a few days into starting effective systemic therapy. The average turnaround time for CTC testing was 5-7 days. Table 1. CTC values in initial baseline blood draw sample Gastrointestinal cancer subtype # of patients Average # of CTCs per 7.5mL blood Median # of CTCs per 7.5mL blood Minimum # of CTCs per 7.5mL blood Maximum # of CTCs per 7.5mL blood Colorectal carcinoma 27 84 3 0 1378 Cholangiocarcinoma 3 8 3 1 23 Pancreatic cancer 3 2 2 0 3 Hepatocellular carcinoma 2 4 5 0 10 Gallbladder cancer 1 6 6 3 8 Appendix cancer 1 2 2 1 3 Anal cancer 1 0 0 0 0 Conclusions: Our study emphasizes that real-time monitoring of CTCs is complementary to traditional tumor burden monitoring via imaging and ctDNA analysis. CTCs offer multiple unique advantages vs ctDNA including early response assessment within days of initiating therapy, a substantial decrease in cost of serial testing, and the ability to assess clinically relevant cell surface protein biomarkers e.g. HER2 and PD-L1. Citation Format: Erin G. Bayer, Brock M. Bartels, Rachel N. Ponting, Areeb Lutfi, Maaz K. Afghan, Arturo B. Ramirez, Pashtoon M. Kasi. Real-time monitoring of CTC number, biomarker expression, and kinetics as prognostic and predictive markers in patients with GI malignancies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7493.

Abstract 7487: Construction of a reliable CTC detection method and validity evaluation of CTC markers in colorectal cancer

Abstract Even in modern times when medical technology has advanced, cancer remains a life-threatening disease, and recurrence and metastasis are major issues in cancer treatment. The characteristics of cancer cells change with progression and treatment. Therefore, timely evaluation on their characteristics and decision on the treatment strategy are required. Circulating tumor cells (CTCs) are released into blood vessels in solid tumors and promote metastasis by circulating in the blood. CTCs are thus attractive targets for liquid biopsy and can provide useful information for selection of appropriate treatment. To obtain reliable results from CTC testing, it is important to prove that the detected cells are derived from cancerous tissue. In molecular tests targeting CTCs, it is still unclear whether the detected cells are derived from cancer tissue displaying genetic mutations. Aiming to establish reliable CTC testing, we have developed a simple method using molecular imaging flow cytometry to detect APC gene mutation together with Cytokeratin and Vimentin, which are epithelial-mesenchymal transition markers. Since, abnormalities in the APC gene are detected in 60-70% of colorectal cancer patients, detecting such mutations provides strong evidence that the detected cells are derived from cancerous tissue. Here, we show that upon evaluation of analytical performance of our method using cells isolated from cancerous tissue, our method showed high concordance rate against APC gene mutation. A clinical proof-of-concept using 5 mL of whole blood from colorectal cancer patients revealed that counts of cells with APC mutation in blood and cells with Cytokeratin and/or Vimentin expression in blood are highly correlated. Our results demonstrated a pathological stage-dependent increase in the count and frequency of Cytokeratin and/or Vimentin positive CTCs. We believe that our study not only establishes a practical approach for identifying CTCs in cancer patients but also provides potential implications in clinical practice, prognostic evaluation, and selection of treatment strategies. Citation Format: Yusuke Takahashi, Yuichi Ijiri, Shiki Fujino, Nakhaei Elnaz, Ayuko Kishimoto, Kentaro Shirai, Shigeki Iwanaga, Masatoshi Yanagida, Ali Asgar Bhagat, Norikatsu Miyoshi. Construction of a reliable CTC detection method and validity evaluation of CTC markers in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7487.

Prognostic Significance of Excision Repair Cross-Complementation Group 1 on Circulating Tumor Cells for Nasopharyngeal Carcinoma.

Liquid biopsy, including the detection of circulating tumor cells (CTCs), has emerged as a promising tool for cancer diagnosis and monitoring. However, the prognostic value of CTCs in nasopharyngeal carcinoma (NPC) remains unclear due to the lack of phenotypic characterization. The expression of Excision Repair Cross-Complementation Group 1 (ERCC1) and CTCs epithelial-mesenchymal transition (EMT) have been associated with treatment efficacy. In this study, we aimed to evaluate the prognostic significance of ERCC1 expression on CTCs and their EMT subtypes before treatment in NPC. We retrospectively analyzed 108 newly diagnosed locally advanced NPC patients who underwent CanPatrol™ CTC testing between November 2018 and November 2021. CTCs were counted and classified into epithelial, epithelial-mesenchymal hybrid, and mesenchymal subtypes. ERCC1 expression was divided into negative and positive groups. Clinical features and survival outcomes were analyzed. The positive rate of CTCs was 92.6% (100/108), with an ERCC1 positivity rate of 74% (74/100). Further analysis of the subtypes showed that positive ERCC1 on mesenchymal CTCs was associated with a later N stage (P = .01). Positive ERCC1 expression was associated with poor overall survival (OS; P = .039) and disease-free survival (DFS; P = .035). Further analysis of subtypes showed that the positive ERCC1 on mesenchymal-type CTCs was associated with poor OS (P = .012) and metastasis-free survival (MFS; P = .001). Our findings suggest that ERCC1 expression on CTCs may serve as a new prognostic marker for NPC patients. Evaluating CTCs subtypes may become an auxiliary tool for personalized and precise treatment.

Effect of circulating tumor cells distribution in treatment naive and treated patients with advance stage breast cancer on disease burden.

e12517 Background: Breast malignancies is a leading cause of cancer-related mortalities and show ascending incidence rate. Despite advancements in our understanding of the disease, its clinical outcome is often dismal. This largely remains owing to the characteristic wide window of relapse, spanning months to decades after primary treatment. Therefore, continuous monitoring of the disease is an offered choice to detect metastatic progression and recurrence. Circulating tumor cells (CTCs) have emerged as a powerful prognostic tool to predict the disease outcome in many epithelial cancers. CTCs are a real-time surrogate biomarker accounting for minimal residual disease (MRD) which is often missed in CT PET scanning. This leads to a progression of metastasis when the patient is often considered as ‘clinically disease free’. Here, we analyzed the presence of CTCs in treatment-naïve and chemo-recipient breast cancer patients. Methods: In this retrospective study on 417 breast cancer patients, CTCs were isolated from 1.5 ml of blood using the Drug Controller General of India (DCGI) approved OncoDiscover CTC test. This platform contains affinity-based magnetic nanoparticles to mediate EpCAM-based CTC isolation. CTCs were detected as CK18+, DAPI+ and CD45- cells based on automated digital imaging platform. Results: 42.6 % (n=178) of patients were clinically at progressive stage (stage II and III) and treatment naïve. On the other hand, 47.4 % of patients received treatment including surgery and chemotherapy. CTCs were not observed in 5.6 % (n=10) of the treatment naïve population while 32% (n=75) of patients who received therapy did not show CTCs. The mean CTC number in treatment naïve patients was 15, while the mean CTC count in patients receiving therapy was drastically reduced to 2. This implied that therapy effectively countered the tumor progression and reduced the shedding of tumor cells in circulation. The distribution of CTC in treatment naïve patients exhibited a bimodal trend centered at values of 10 and 50, suggesting two distinct populations of patients with respect to CTC count. CTC count did not show any correlation with the age in both population groups. Surprisingly CTC count in younger patients (20-50 years) was 50% higher compared to the older population (50-75 years). Conclusions: The presence of CTCs in treatment naïve, progressive breast cancer patients indicated the biologically aggravated disease. Although therapeutic intervention drastically reduced the CTC burden, their presence in a large population was suggestive of an MRD and the likelihood of recurrence after discontinuation of therapy. A distinct pattern of CTC occurrences in Tx naïve and Tx recipient patient suggested that CTCs can be an important clinical indicator to monitor the therapy response, progression and residual disease.

Circulating tumor cells (CTCs) detection and isolation in different subtypes of early-stage breast cancer patients from Bangladesh.

e12529 Background: Breast cancer is a highly heterogeneous pathophysiology characterized by poor outcomes. Due to the increasing incidence and disease progression rates and undefined relapse periods, reliable disease monitoring is a challenge and has remained an unmet need. Advancements in liquid biopsy have significantly enhanced our understanding of clinical oncology. CTC-based liquid biopsy is emerging as a reliable prognostic tool to predict various clinical indicators. Although extensively investigated in metastatic breast cancers, little is known about CTCs in early-stage breast cancers. CTCs with respect to different molecular subtypes of breast cancer in early-stage breast cancer patients is evaluated. Methods: In this prospective clinical trial (CMC 59.27.0000.013.19 PG.009.2022/262) 40 early-stage patients with luminal (A +B, 33.33%), HER 2 positive (12.8%), triple-negative (12.8%) and undetermined (41.07%) subtype were recruited. CTCs were isolated in 1.5 ml blood using the Drug Controller General of India approved OncoDiscover CTC test. This platform contains affinity-based magnetic nanoparticles to mediate EpCAM-based CTC isolation. CTCs were detected as CK18+, DAPI+, and CD45- cells using a fluorescence detection-based automated digital imaging platform. Results: CTCs were detected in 60 % of patients with a mean CTC count of 1 cell / 1.5ml blood. Among total positive patients, the luminal subtype was the least positive (46 %), followed by TNBC (60 %) and undetermined (62.5 %) subgroup, while all HER2-positive patients showed the presence of CTCs. Besides individual cells, CTC clusters were detected in 12.5 % of patients and they were equally distributed in luminal and HER2-positive subpopulations. When analyzed on the scale of tumor grade, grade I patients did not show the presence of CTC, while 58.33 % of grade II patients had ≥ 1 CTC. All grade III patients showed the presence of ≥ 1 CTC. CTC count was high among CTC-positive grade II patients (average 2 CTCs) and correlated well with the presence of CTC clusters in these patients. Patients who had surgical intervention had a low CTC burden compared to patients who did not have a surgical resection. 75 % of treatment naïve patients showed the presence of CTC while 58 % of patients receiving chemotherapy alone showed the presence of 1 CTC. 50 % of patients who had surgery followed by CT+RT showed the presence of 1 CTC. Conclusions: The presence of CTCs may suggest for biological progression of disease in early-stage BC patients. CTC detected in all HER2-positive patients suggested the high shedding nature of these tumors, which correlates well with their reported migratory tendency. The presence of CTCs did not show a clear correlation with the treatment regimen. However, this data is based on a single time point and needs longitudinal correlation with CTC on a larger sample size. Clinical trial information: 59.27.0000.013.19 PG.009.2022/262 .

Feasibility study of expressing epcam + /vimentin + CTC in prostate cancer diagnosis.

Prostate cancer (PCa) is one of the most common malignancies in men and one of the leading causes of cancer-related deaths; circulating tumor cells (CTC) are malignant cells that have broken off from original tumor or metastatic sites and extravasated into the blood vessels either naturally or maybe as a consequence of surgical procedures. This study aims to explore the feasibility of liquid biopsy technique to diagnose prostate cancer. We constructed an assay platform integrating magnetic separation and fluorescence in situ hybridization (FISH) to effectively capture prostate cancer CTCs and evaluate the distribution between healthy volunteers and prostate cancer patients, respectively. There was a significant difference in the number of CTCs between the healthy population and prostate cancer patients (P < 0.001). The results of the study showed that the CTCs capture identification system has good sensitivity and specificity in identifying prostate cancer patients. The CTCs test allows us to accurately identify patients who are at high risk for prostate cancer, allowing for early intervention and treating patients effectively.

Abstract P1-05-23: OPTIMIZING THE DIAGNOSIS OF LEPTOMENINGEAL METASTASES IN BREAST CANCER PATIENTS BY CIRCULATING TUMOR CELLS AND CIRCULATING TUMOR DNA

Abstract Introduction: Leptomeningeal metastases (LM) occur in approximately 5% of patients with breast cancer, negatively impacting their prognosis. Reliable diagnosis of LM is challenging, but timely diagnosis could lead to more treatment options and consequently a better prognosis. Circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs) in cerebrospinal fluid (CSF) are potential methods to improve the diagnosis of LM. In this prospective study we compared cytology (current gold standard for CSF analysis in diagnosing LM) with CTC enumeration and ctDNA detection in CSF for diagnosing LM in patients with breast cancer. Methods: In a prospective study in patients with clinical suspicion of LM who were planned to undergo a diagnostic lumbar puncture (LP) for CSF cytology the presence of CTCs/ctDNA in CSF was compared to the presence of tumor cells in CSF by cytology. CTC enumeration was performed using the FDA approved CellSearch System (Menarini). The detection of ctDNA was performed using the mFAST-Seqs method, which detects aneuploidy throughout the genome by selective amplification of long interspaced nuclear element sequences (LINE-1 elements). In a control group consisting of 15 patients with a hematologic malignancy with suspicion of LM, 2 cases had 1 CTC in their CSF, therefore the cutoff for a positive CTC test was set at 2 CTCs. Results: In total 80 patients with breast cancer were included from January 2016 till January 2022. In 16 out of 80 patients CSF cytology was positive of whom 14 (87.5%) had a positive CTC test. From the 2 patients in which the CTC test was negative, one patient had 1 CTC detected in CSF and for both patients a lower volume of CSF was available for the CTC test compared to the volume used for cytology. In 2 of the 16 cytology positive patients, who underwent a second LP, CTCs were positive in the CSF from the first LP, while cytology was only positive in the CSF of the second LP. Of the 64 patients with a negative cytology, 7 (10.9%) patients had a positive CTC test. Two of these 7 patients had typical symptoms of LM, a suspicious MRI and a course of disease matching with LM according to the neuro-oncologist. Overall survival (OS) was shorter in patients with a positive CTC test compared to patients with a negative test (median OS: 19.2 months vs. 3.4 months, log-rank test, p=0.007). When only patients with a negative cytology were included in this analysis, OS remains significantly shorter in the group with a positive CTC test (n=7) compared to the group with the negative CTCs (n=57) (median OS 19.3 months vs. 3.5 months, long-rank test, p=0.003). Additionally, ctDNA detection in CSF was compared to CSF cytology in 51 patients, of which 11 had a positive CSF cytology result. For 8 of these 11 patients ctDNA was detected (72.2%) while 1 of the 40 patients with a negative cytology had ctDNA. Discussion: Both CTC enumeration and ctDNA detection can be used to detect LM in CSF. CTC detection could even improve timely diagnosis of LM in patients with breast cancer. However, the added value of ctDNA seems less evident. Citation Format: Elisabeth M Jongbloed, Lindsay Angus, Martin J van den Bent, Joost LM Jongen, John WM Martens, Ngoc M Van, Vanja de Weerd, Carolien HM van Deurzen, Jaco Kraan, Saskia M Wilting, Agnes Jager. OPTIMIZING THE DIAGNOSIS OF LEPTOMENINGEAL METASTASES IN BREAST CANCER PATIENTS BY CIRCULATING TUMOR CELLS AND CIRCULATING TUMOR DNA [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 P1-05-23.

Diagnostic and prognostic value of circulating tumor cells in Indian women with suspected ovarian cancer.

"Liquid biopsy," where body fluids are screened for biomarkers, is gathering substantial research. We aimed to examine women with suspected ovarian cancer for the presence of circulating tumor cells (CTCs) and study its role in prediction of chemoresistance and survival. Magnetic powder labeled monoclonal antibodies for epithelial cell adhesion molecule (EpCAM), mucin 1 cell surface associated, mucin 16 cell surface associated, or carbohydrate antigen 125 (CA125), were prepared according to the manufacturer's protocol. Expression of three ovarian cancer related genes was detected in CTCs using multiplex reverse transcriptase-polymerase chain reaction. CTCs and serum CA125 were measured in 100 patients with suspected ovarian cancer. Correlations with clinicopathological parameters and treatment were analyzed. CTCs were detected in 18/70 (25.7%) of women with malignancy compared to 0/30 (0.0%) in those with benign gynecologic diseases (P = 0.001). The sensitivity and specificity of the CTC test for predicting a malignant histology in pelvic masses were 27.7% (95% CI: 16.3%, 37.7%) and 100% (95% CI: 85.8%, 100%), respectively. The number of CTCs correlated with stage of ovarian cancer (P = 0.030). The presence of EpCAM + CTC at primary diagnosis in ovarian cancer was found to be an independent predictor of a poor progression free survival (HR, 3.3; 95% CI, 1.3-8.4; P = 0.010), overall survival (HR, 2.6; 95% CI,1.1-5.6; P = 0.019), and resistance to chemotherapy (OR 8.6; 95% CI, 1.8-43.7; P = 0.009). Expression of EpCAM + CTC in ovarian cancer predicts platinum resistance and poor prognosis. This information could be further used in investigating anti-EpCAM-targeted therapies in ovarian cancer.

Clinical Relevancy of Circulating Tumor Cells in Breast Cancer: Epithelial or Mesenchymal Characteristics, Single Cells or Clusters?

Metastatic breast cancer (MBC) is typically an incurable disease with high mortality rates; thus, early identification of metastatic features and disease recurrence through precise biomarkers is crucial. Circulating tumor cells (CTCs) consisting of heterogeneous subpopulations with different morphology and genetic, epigenetic, and gene expression profiles represent promising candidate biomarkers for metastatic potential. The experimentally verified role of epithelial-to-mesenchymal transition in cancer dissemination has not been clearly described in BC patients, but the stemness features of CTCs strongly contributes to metastatic potency. Single CTCs have been shown to be protected in the bloodstream against recognition by the immune system through impaired interactions with T lymphocytes and NK cells, while associations of heterotypic CTC clusters with platelets, leucocytes, neutrophils, tumor-associated macrophages, and fibroblasts improve their tumorigenic behavior. In addition to single CTC and CTC cluster characteristics, we reviewed CTC evaluation methods and clinical studies in early and metastatic BCs. The variable CTC tests were developed based on specific principles and strategies. However, CTC count and the presence of CTC clusters were shown to be most clinically relevant in existing clinical trials. Despite the known progress in CTC research and sampling of BC patients, implementation of CTCs and CTC clusters in routine diagnostic and treatment strategies still requires improvement in detection sensitivity and precise molecular characterizations, focused predominantly on the role of CTC clusters for their higher metastatic potency.

Detection and surveillance of circulating tumor cells in osteosarcoma for predicting therapy response and prognosis.

Objective:Osteosarcoma (OS) is an aggressive, highly metastatic, relatively drug-resistant bone tumor with poor long-term survival rates. The presence and persistence of circulating tumor cells (CTCs) in the peripheral blood are believed to be associated with treatment inefficiency and distant metastases. A blood-based CTC test is thus greatly needed for monitoring disease progression and predicting clinical outcomes. However, traditional methods cannot detect CTCs from tumors of mesenchymal origin such as OS, and research on CTC detection in mesenchymal tumors has been hindered for years.Methods:In this study, we developed a CTC test based on hexokinase 2, a metabolic function-associated marker, for the detection and surveillance of OS CTCs, and subsequently explored its clinical value. Twelve patients with OS were enrolled as the training cohort for serial CTC tests. Dynamic CTC counting, in combination with therapy evaluation and post-treatment follow-up, was used to establish a model for predicting post-chemotherapy evaluation and disease-free survival, and the model was further validated with a cohort of 8 patients with OS.Results:Two dynamic CTC number patterns were identified, and the resulting predictive model exhibited 92% consistency with the clinical outcomes. This model suggested that a single CTC test has similar predictive power to serial CTC analysis. In the validation cohort, the single CTC test exhibited 100% and 87.5% consistency with therapy response and disease-free survival, respectively.Conclusions:Our non-invasive test for detection and surveillance of CTCs enables accurate prediction of therapy efficiency and prognosis, and may be clinically valuable for avoiding inefficient therapy and prolonging survival.

Partial advances in the diagnosis and treatment of gastrointestinal cancer.

Partial advances in the diagnosis and treatment of gastrointestinal cancer.

Predictive value of circulating tumor cells FTH1 gene on the efficacy of neoadjuvant chemotherapy in non-metastatic breast cancer.

e12599 Background: Based on epithelial-mesenchymal transition (EMT) markers, circulating tumor cells (CTCs) can be classified into two main subtypes: epithelial CTCs(E-CTCs) and EMT-CTCs. CTCs with high expression of the FTH1 gene (HEF) may be related to poor efficacy of neoadjuvant chemotherapy (NAC). The relation between the CTCs with HEF and pathological complete response (pCR) after NAC is unclear. Methods: 83 non-metastatic breast cancer patients who completed NAC and surgery were enrolled in this study, and each patient had at least one result of CTC test before surgery. Expression of the FTH1 gene on CTCs was achieved by multiplex RNA in situ hybridization. Mann-Whitney U tests was conducted to examine the differences between the non-pCR and pCR groups in the numbers of total CTCs, E-CTCs and EMT-CTCs with HEF. Predictive ability of total and subtypes of CTCs with HEF to pCR rate was compared by the area under receiver operating characteristic curves (AUC). Univariate and multivariate binary logistic regression were used to select the variable and construct the predictive model. Results: Among 64 patients who underwent CTC test before NAC, CTCs were found in 52 patients, of which 20 patients achieved pCR. There was no significant difference in the numbers of total CTCs (9.9±10.9 vs 7.9±7.1, p=0.828) and E-CTCs with HEF (0.6±0.9 vs 0.3±0.7, p=0.256). Compared with the patients in the non-pCR group, those in pCR group had fewer EMT-CTCs with HEF (2.5±5.1 vs 1.0±2.5, p&lt;0.01). 56 patients had CTC test after NAC, CTCs were detected in 53 patients, of which 18 patients gained pCR. There was no significantly difference in the numbers of total CTCs (11.7±13.8 vs 7.1±6.2, p=0.386), E-CTCs with HEF (0.5±1.0 vs 0.3±0.6, p=0.699) and EMT-CTCs with HEF (3.6±7.9 vs 1.3±3.1, p=0.323) between the two groups. In patients with CTC tests before NAC, EMT-CTCs with HEF have the highest predictive ability (AUC=0.718, 95%CI=0.567-0.869) than total CTCs (AUC=0.518, 95%CI=0.359-0.677) and E-CTCs with HEF (AUC=0.575, 95%CI=0.417-0.733). Univariate binary logistic regression analysis showed that HR status (OR=4.667, 95%CI= 1.177-18.506), HER2 status (OR=0.059, 95%CI=0.014-0.255) and EMT-CTCs with HEF status (&lt;1 vs ≥1) (OR=8.800, 95%CI=2.336-33.152) were significantly related with the pCR rate. These variables remain significant in the multivariate logistic analysis. The AUC value of the pCR rate prediction model constructed based on the multivariate binary logistic regression results was significantly higher than the model combining HR status and HER2 status (AUC=0.930, 95%CI=0. 861-1.000 vs AUC=0.866, 95%CI=0.765-0.966; p&lt;0.05), based on Delong test. Conclusions: EMT-CTCs with HEF before NAC has the predictive ability for the rate of pCR in patients with non-metastatic breast cancer. This will help clinicians personalize the treatment strategy for patients to achieve better survival.

Guiding Value of Circulating Tumor Cells for Preoperative Transcatheter Arterial Embolization in Solitary Large Hepatocellular Carcinoma: A Single-Center Retrospective Clinical Study.

BackgroundLarge hepatocellular carcinoma (LHCC) is highly malignant and prone to recurrence, leading to a poor long-term prognosis for patients. There is an urgent need for measures to intervene in postoperative recurrence. Preoperative Transcatheter Arterial Embolization (TACE) is an effective treatment. However, there is a lack of reliable preoperative indicators to guide the application of preoperative TACE. We, therefore, investigated whether the preoperative status of circulating tumor cells (CTCs) could be used to guide preoperative TACE for HCC treatment.MethodsThis study recruited 361 HCC patients and compared recurrence-free survival (RFS) and overall survival (OS) in patients treated with TACE prior to surgery and those not treated with TACE. Patients were divided into CTC-positive group and CTC-negative group according to CTC status, and the effect of preoperative TACE on RFS and OS was compared in each subgroup.ResultsIn CTC-positive patients, preoperative TACE reduces early recurrence and improves long-term survival. However, HCC patients did not benefit from preoperative TACE for the overall population and CTC-negative patients.ConclusionsPreoperative CTC testing is a reliable indicator of whether HCC patients received TACE preoperatively. CTC positivity was associated with early tumor recurrence, and preoperative TACE could reduce early recurrence and long-term prognosis in CTC-positive patients.

Circulating Tumor Cell Identification Based on Deep Learning.

As a major reason for tumor metastasis, circulating tumor cell (CTC) is one of the critical biomarkers for cancer diagnosis and prognosis. On the one hand, CTC count is closely related to the prognosis of tumor patients; on the other hand, as a simple blood test with the advantages of safety, low cost and repeatability, CTC test has an important reference value in determining clinical results and studying the mechanism of drug resistance. However, the determination of CTC usually requires a big effort from pathologist and is also error-prone due to inexperience and fatigue. In this study, we developed a novel convolutional neural network (CNN) method to automatically detect CTCs in patients’ peripheral blood based on immunofluorescence in situ hybridization (imFISH) images. We collected the peripheral blood of 776 patients from Chifeng Municipal Hospital in China, and then used Cyttel to delete leukocytes and enrich CTCs. CTCs were identified by imFISH with CD45+, DAPI+ immunofluorescence staining and chromosome 8 centromeric probe (CEP8+). The sensitivity and specificity based on traditional CNN prediction were 95.3% and 91.7% respectively, and the sensitivity and specificity based on transfer learning were 97.2% and 94.0% respectively. The traditional CNN model and transfer learning method introduced in this paper can detect CTCs with high sensitivity, which has a certain clinical reference value for judging prognosis and diagnosing metastasis.

Tend and forecast of Global Market instruments for laboratory Diagnostic

In-vitro diagnostics is the process of diagnosing various diseases using test kits and analyzers. The in vitro diagnostics (IVD) market is growing at a steady pace as awareness of various diseases grows and the government initiates various plans to reduce the incidence. Most IVD tests are performed in medical and clinical laboratories, but the advent of point-of-care (POC) technology has enabled testing in the presence of the patient and even at home. This technological advancement is driving the growth of the market over the forecast period. Market growth over the forecast period is expected to be driven by factors such as the increase in lifestyle-related diseases, the growing adoption of POC testing, the growing demand for biomarker-based testing, and demand from developing countries. Factors hindering the growth of the market are the increase in sales of counterfeit glucose test strips and strict regulatory approval procedures. The regulatory scenario is of concern as there is a wide range of tools and analyzes that range from Class I to Class III devices. Recently, there have been changes in legislation in the European Region, which has become a serious problem for market participants. There is an emerging trend in the market towards non-invasive prenatal testing, next generation sequencing (NGS), fluid biopsy and circulating tumor cell testing. Markets in India, Brazil and China are expected to grow at a rapid pace during the forecast period due to the high prevalence of infectious diseases in these countries. The purpose of this study is to assess the current global market for medical devices for in vitro diagnosis, identify the main factors and drivers influencing the growth and prospects of this market in the future. To conduct this study, as materials and methods, we used the analysis of these indicators of expert assessments published by specialists in Russian and international journals specializing in the implementation, promotion and development of laboratory diagnostic products. The results of the study determine the key promising markets for medical devices for in vitro diagnostics, as well as the correct vector for the development of the medical industry. In the final part, the main regions with the highest development potential over the next 10 years were identified, and indicators of the dynamics of the global market for medical instruments over the past 5-9 years were determined.