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Combined ctDNA and CTC analysis improves MRD detection and cancer progression monitoring. Publications 3 June 2024 ASCO 2024: Association of complementing ctDNA and CTCs load on stable and progressive disease in treated patients. Complementary ctDNA and CTC biomarkers reveal minimal residual disease and predict cancer progression after curative-intent treatment. Background: Post curative-intent surgery and therapy, the presence of circulating tumor DNA (ctDNA) load represents minimal residual disease (MRD). Conversely, the presence of circulating tumor cells (CTCs) in stage I–II cancer or even in disease-free survival (DFS) patients indicates occult cellular residual disease (CRD) with undetectable micrometastasis. These complementary biomarkers in patients undergoing treatment act as indicators of non-responsiveness, suggesting the need for treatment modifications. Methods: Retrospectively, we monitored a cohort of 46 cancer patients for MRD using ctDNA and CTCs who were treated or undergoing treatment (e.g., lung, breast, colon, and head and neck cancer; n = 14, 7, 6, and 4, respectively). The OncoMonitor test detected single nucleotide variations (SNVs), small insertions and deletions (INDELs), copy number variations (CNVs), and translocations (fusions). Libraries were prepared using a hybridization-capture method covering 1000 targets with a mean sequencing depth of 5000× on the Illumina NextSeq 2000 in paired-end mode (150 × 2). Variant calling was performed using a proprietary bioinformatics pipeline, iCare. CTCs were isolated using the OncoDiscover platform, which possesses an anti-EpCAM antibody-based immunomagnetic system per 1.5 mL of blood. CTCs were confirmed using CK18+, PD-L1, and CD45 markers with a motorized fluorescence microscope. Results: From ctDNA analysis, 47.82% (n = 22) of patients were identified with at least one actionable genomic finding. Among these, 13.04% (n = 6) of patients showed EGFR driver mutations. Additionally, 19.56% (n = 9) of patients were identified with either EGFR driver, KRAS, or PI3K passenger mutations, while 4.34% (n = 2) were identified with ALK–EML4 fusion. The average ctDNA load obtained in patients with progressive disease (n = 26) was 8.2 molecules per 1 mL of plasma. At least one CTC was detected in 61.53% (n = 16) of progressive disease patients, with the highest count of four CTCs identified in 7.69% (n = 2) of patients. Only 30% (n = 6) of patients with stable disease were identified with at least one genomic finding from a total of 20 patients upon ctDNA analysis, with an average ctDNA load of 2.2 molecules per 1 mL of plasma. Patients with clinically progressive disease showed ctDNA load approximately fourfold higher than those with stable disease during treatment. No patients with stable disease were identified with four CTCs, as opposed to 7.69% in the progressive disease cohort during treatment. Conclusions: We observed that ctDNA and CTCs complement MRD status even after curative-intent surgery and therapy, with the potential to identify patients likely to experience disease progression. Our findings strongly indicate a positive correlation between ctDNA load, the number of detected CTCs, and disease progression based on radiological findings. These biomarkers can support practical clinical decision-making. Further studies are necessary to validate these findings and improve follow-up strategies for better clinical outcomes. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Rick Kamble Cofounder & Director Actorius Innovations and Research Pvt. Ltd. Rick Kamble Cofounder & Director LinkedIn Qualification Master of Business Administration University of Phoenix, US Bachelor of Engineering Nagpur University, India Professional Summary Rick is an early investor in Actorius Innovations and Research (AIR) and chairman of the board. He is a serial entrepreneur based our of Los Angeles, US with expertise in Healthcare and Information Technology solutions. Rick is also the CEO and President of HealthNspire solutions, a HealthCare Technology solution provider focused on managed care software solutions. He is also the co-founder and CTO of Mediccio, a company dedicated to provide healthcare billing solutions using NLP. In the past, Rick has served as Senior VP of Product Strategies at AssistMed, a company dedicated to the application of Healthcare IT solutions. He has held leadership positions at Lakeside Medical Group, Jacobs Engineering, Farmers Insurance, L.A. CARE Health Plan, Cardinal Health, Countrywide Home Lo Want to be a part of this dynamic team? Visit our Careers section to explore current opportunities that match your interests and expertise. Apply Now

CTC and PD-L1 analysis helps detect MRD and guide therapy in colorectal cancer. Publications 3 June 2024 ASCO 2024: Measure of minimal residual burden on CTCs with over-expression of PD-L1 as a dynamic biomarker in patients with colorectal cancer. CTC detection with PD-L1 expression in colorectal cancer reveals minimal residual disease and supports personalized treatment strategies. Background In stage III colorectal cancer (CRC) patients, the extent of oxaliplatin-based adjuvant therapy remains uncertain. Approximately 25–50% of stage II–III CRC patients develop recurrence and metastasis even after comprehensive treatment, largely attributed to occult disease and minimal residual disease (MRD). Circulating tumor cells (CTCs) represent a bio-mechanistic source of extravasation leading to micro-metastatic disease. CRC patients receiving reduced adjuvant therapy (3–6 months) are known to exhibit increased CTC counts and positivity rates due to the emergence of resistant clones. Assays that detect CTCs and the expression of programmed death-ligand 1 (PD-L1) as a dynamic biomarker simultaneously have significant clinical implications, particularly when tissue biopsy samples are inadequate to identify molecular targets for immune checkpoint inhibitor (ICI) therapy. Methods In a retrospective study, 182 CRC patients were analyzed for the presence and distribution of CTCs at baseline and across follow-ups (0–4 follow-ups). Peripheral blood (1.5 ml) samples were analyzed using the CDSCO-approved OncoDiscover platform, which consists of a multifunctional magneto-nanosystem mediated by anti-epithelial cellular adhesion molecule (EpCAM) antibodies. CTCs were evaluated in patients with early-stage disease (pre- and post-treatment), progressive disease, disease-free status (DFS), and metastasis. Isolated cells were immunostained to detect CK18+, CD45-, DAPI+, and PD-L1+ expression. PD-L1 expression on CTCs was validated by analyzing the linear intensity gradients of fluorescence signals. CTCs were classified as PD-L1 negative when weak or no fluorescence signal was detected and PD-L1 positive when strong fluorescence signals were observed using automated image acquisition on a Zeiss fluorescence microscope. Results Among the cohort of 182 CRC patient samples, 128 (70.3%) showed the presence of CTCs. A fluorescence intensity-based assay was developed to evaluate PD-L1 expression as a robust functional biomarker for molecular characterization of CTCs. The distribution of CTCs ranged from 1 to 9 cells. The mean fluorescence intensity value and cut-off for PD-L1 expression in CTCs was approximately 1.02. Notably, 54 patients (42.2%) with CTCs showed positive PD-L1 expression. CTC-positive patients with PD-L1 expression were observed across all stages, including early-stage disease, progressive disease, and metastasis. Patients without detectable CTCs (n = 54, 29.7%) either had clinically stable disease or were in DFS with no radiographic evidence of disease. Conclusions PD-L1 overexpression on CTCs represents a dynamic blood-based biomarker indicating disease progression even in patients with DFS status. Enumeration of CTCs along with assessment of PD-L1 expression may enable more individualized treatment strategies for CRC patients and support better monitoring of disease progression and therapeutic response. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Actorius Innovations AACR 2026 Publications Publications 17 March 2026 AACR 2026: Depletion of circulating tumor cells using an automated device using non-hemolytic affinity-based substrates Actorius Innovations presents accepted research abstracts at the AACR Annual Meeting 2026, highlighting advances in cancer diagnostics, therapeutics and liquid biopsy. Abstract Background While 90% of cancer deaths are associated with metastasis, it is imperative to monitor early-stage cancer patients for the presence of systemic disease to improve overall survival (OS) and progression-free survival (PFS). Despite complete remission, up to 25–50% of colorectal cancer (CRC) stage II–III and early breast cancer cases are known to relapse. Furthermore, the existence of microtumors often remains undetected by radio-imaging tools due to their limited detection thresholds. Following curative-intent therapies, minimal residual cellular disease (MRCD) may persist and is often represented by circulating tumour cells (CTCs). These cells are known for their ability to extravasate and invade distant sites from the primary tumor. They can also evade immune surveillance. Therefore, there is a need to design safer extracorporeal devices for the capture and depletion of CTCs, particularly those overexpressing PD-L1. In this study, we designed an automated device to capture and remove CTCs from whole blood. Methods We developed an automated microprocessor-operated fluidic device, OncoMetastat, equipped with cartridges for blood and reagent tubes, along with a 3D-printed biocompatible spiral channel. The controller unit powers peristaltic pumps that circulate blood through the spiral channel (96 mm diameter × 6 mm height). The system incorporates 2 mm glass beads conjugated with antibodies and transferrin. Additionally, four vibrators provide micro-stirring to enhance CTC capture from 5–10 mL of patient blood samples (n = 54). White blood cell (WBC) count, hemolysis, and protein binding were measured. The beads were scanned for CTCs using markers CK18⁺, DAPI⁺, and CD45⁻ through an automated imaging system and compared with the OncoDiscover CTC enumeration platform approved by CDSCO India. We analyzed true positives, false negatives, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy. Results Retrospectively, blood samples from 54 pan-cancer patients—including breast, colorectal, prostate, and lung cancer—were analyzed to capture and deplete CTCs. The OncoMetastat platform demonstrated a capture efficiency of over 90% when compared with the OncoDiscover platform. Automated scanning achieved 100% efficiency in CTC imaging compared with manual imaging. Leukocyte adhesion was low with anti-EpCAM and transferrin-coated glass beads (2 ± 1 WBCs per sample, n = 54). WBC counts showed cancer-type-specific trends (mean WBC count/mL: 4.9 × 10⁶ for breast cancer, 3.9 × 10⁶ for rectal cancer, and 3.5 × 10⁶ for prostate cancer), representing a 40% decrease compared with healthy controls (mean 6.9 × 10⁶ WBCs/mL). Clinically insignificant hemolysis (<1%) and minimal protein binding (~1.5%) were observed. Vibration-assisted operation enhanced CTC sequestration, achieving more than 90% cell capture efficiency. The platform demonstrated sensitivity of 94.4%, specificity of 92.9%, PPV of 94.4%, NPV of 92.9%, and overall accuracy of 93.8% for CTC capture. Conclusion This study demonstrates efficient and specific depletion of CTCs using the automated device. The platform shows potential as an extracorporeal system capable of removing CTCs from whole blood, thereby offering a promising strategy to enhance cancer therapy outcomes. View Publication Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

PD-L1 on CTCs enables monitoring, recurrence tracking, and minimal residual disease. Publications 3 June 2025 Circulating tumor cells and clusters exhibiting expression of PD-L1 in colorectal patients. High prevalence of PD-L1–positive circulating tumor cells in colorectal cancer highlights their role in minimal residual disease and recurrence monitoring. Background The role of circulating tumor cells (CTCs) has been well established in predicting survival in metastatic settings, particularly in breast, colorectal, and prostate cancers. However, their clinical utility has been limited due to high costs, variability in sensitivity and accuracy, and the use of cutoff-based interpretations. The biological significance of CTCs—from extravasation and invasion to their contribution to tumor microenvironment dynamics and tumor burden—suggests greater clinical relevance than is currently applied in practice. Their role in monitoring minimal cellular residual disease (MCRD), especially in early-stage cancers post-surgery, remains underexplored, including decisions regarding therapy duration in diseases such as colorectal cancer and longitudinal monitoring for recurrence. Dynamic PD-L1 expression on CTCs may indicate incomplete tumor resection or treatment response and may also reflect cellular dormancy in circulation, potentially enabling immune evasion. In this study, we report the expression of PD-L1 on CTCs and CTC clusters in colorectal cancer patients. Methods We retrospectively analyzed 666 colorectal cancer patients (63.06% male and 36.94% female), spanning early- to late-stage disease, for the presence of CTCs with and without PD-L1 expression, as well as CTC clusters. CTCs were detected using the CDSCO-approved OncoDiscover platform in 1.5 mL of peripheral blood. Cells were classified as CTCs if they were EpCAM⁺, CK18⁺, DAPI⁺, and CD45⁻, and were identified using an automated Zeiss microscope system. Results At baseline analysis, 74.25% (n = 591) of patients had ≥1 CTC per 1.5 mL of blood. CTC counts ranged from 1 to 20 cells. Among patients with detectable CTCs, 74.62% (n = 441) exhibited PD-L1 expression. The highest proportion of CTCs (~25.86%, n = 352) was observed in the 61–70 years age group. CTC clusters were detected in 13.00% (n = 156) of patients, and notably, more clusters were observed during follow-up compared with baseline. The mean CTC count (including clusters) was 1.71, while the mean PD-L1–positive CTC count was 1.02. Conclusions PD-L1 expression on CTCs may contribute to their ability to persist in circulation through immune evasion, potentially enabling dormancy via surface protein overexpression that helps them avoid elimination by immune T cells. The CTC–PD-L1 assay shows strong potential for patient surveillance both before and after treatment in assessing minimal cellular residual disease. Further clinical studies in this direction are strongly warranted. View Publication Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

AI-based image analysis rapidly profiles CTC morphology and biomarker expression. Publications 16 September 2025 Profiling of PD-L1 and HER2 over expression on cancer cells using AI based macro-driven automation AI-based image analysis rapidly profiles circulating tumor cells, quantifying morphology and biomarkers like PD-L1 and HER2 for cancer research. Abstract Background Extravasation, invasion, epithelial-to-mesenchymal transitions, metastasis progression, immune evasion, and therapeutic resistance are driven by phenotypic alterations in cancer cells. Assessing cell morphology, stiffness, and deformability is therefore crucial. The expression of PD-L1, HER2, EGFR, and cytokeratins (CKs) serves as key phenotypic biomarkers for precision oncology. We developed an AI-based image analysis tool that rapidly captures these transitions in cell assays, including specific protein biomarkers expressed on circulating tumor cells (CTCs). Methods We extended an ImageJ macro to enable rapid and reproducible extraction of biophysical parameters. The macro processes .lif, .nd2, and .czi file formats, using DAPI for nuclear segmentation and fluorophore-conjugated antibodies to delineate cytoplasmic boundaries. We evaluated automatic channel detection, intensity normalization, Otsu thresholding, and per-cell quantification of parameters such as surface area, circularity index (CI), and mean fluorescence intensity. Violin plots illustrated temporal variations in CI across A549 and MCF7 cells. Validation was conducted on CTCs isolated from cancer patient samples (n = 100) for PD-L1 and HER2 expression. Results The macro reduced image processing time from 7 minutes to 3 seconds per sample. A549 cells showed higher and more consistent CI values across all time points, while MCF7 cells demonstrated lower CI with greater variability, particularly at 24 and 72 hours. Quantitative measurements of PD-L1 and HER2 expression showed 100% concordance between the ImageJ macro and Zeiss software outputs, confirming analytical accuracy. CK18 intensity (~60–400) and PD-L1 (~20–50) levels measured by both platforms validated the macro’s ability to detect a wide range of marker expression in CTC subsets. CTCs exhibited higher CI values and greater morphological heterogeneity, consistent with their invasive phenotype. Conclusions We present an AI-driven macro that quantifies the biophysical characteristics of cancer cells, enabling precise phenotypic profiling, including circularity index, proliferation rates, and overexpression of biomarkers such as PD-L1 and HER2 in both cultured cell lines and patient-derived CTCs. View Publication Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Real-Time Therapy Response Monitoring Using Surface Biomarkers on Circulating Tumor Cells Publications 27 January 2026 Manuscript: Real-Time Therapy Response Monitoring Using Surface Biomarkers on Circulating Tumor Cells Circulating tumor cells (CTCs), cancer cells shed from primary tumors into the bloodstream, are emerging as dynamic, non-invasive biomarkers for real-time cancer monitoring, especially when tissue biopsies are inaccessible or inadequate... Simple summary Circulating tumor cells (CTCs), which are cancer cells shed from primary tumors into the bloodstream, are emerging as dynamic, non-invasive biomarkers for real-time cancer monitoring, especially when tissue biopsies are inaccessible or inadequate. Unlike static tissue samples, CTCs allow repeated assessments that track tumor evolution, therapeutic response, and minimal residual disease. Hence, CTCs offer a minimally invasive, real-time alternative to tissue biopsies for cancer monitoring, particularly through surface protein biomarkers like PD-L1, HER2, and EGFR. As detection technologies improve and the clinical relevance of CTCs continues to be established, CTC profiling is poised to significantly influence the future of precision oncology. Abstract Circulating tumor cells (CTCs) are shed from the primary tumor into the bloodstream and represent dynamic molecular biomarkers for monitoring the progression of cancer. While profiling tumor tissues with overexpression of cell surface markers, such as PD-L1 or HER2, is standard in guiding therapy, tissue samples are often inaccessible and inadequate, especially post-surgery or in cases of recurrence. Emerging clinical evidence indicates that CTC counts and biomarker surface expression can predict prognosis and therapeutic resistance more accurately than imaging or tissue-based approaches. Recent advancements in CTC detection methods, based on physical properties or surface markers (e.g., EpCAM), coupled with next-generation sequencing (NGS), have enabled the isolation of these rare cells and their molecular characterization. Consequently, CTCs provide a real-time alternative, enabling repeated, longitudinal assessment of tumor phenotype and therapeutic response. This review emphasizes the translational potential of surface protein biomarkers on CTCs for profiling, namely PD-L1, HER2, and EGFR, as a clinically actionable approach to stratify patients, guide immunotherapy decisions, and monitor minimal residual disease (MRD), especially when longitudinal tissue biopsies are not feasible. View Manuscript Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Dr. Jayant Khandare Cofounder, MD & CSO Actorius Innovations and Research Pvt. Ltd. Dr. Jayant Khandare Cofounder, MD & CSO LinkedIn Qualification Master of Pharmacy (M. Pharm.) University of Mumbai | 1995 Ph.D. (Chemical Engineering) National Chemical Laboratory (NCL), Pune | 2003 Alexander von Humboldt (AvH) Experienced Researcher. Humboldt Foundation, Germany | 2008 Fellow of Royal Society of Chemistry (FRSC). Royal Society of Chemistry, UK | 2014 25+ US Patents, 100+ Peer Reviewed Scientific Articles Postdoctoral stints at Wayne State University (USA), Rutgers University (USA) and Freie Universität Berlin (Germany) in the area of targeted drug delivery for cancer. Research interests at the interface of macromolecular chemistry, biopolymer sciences, cancer biology, and cellular imaging. Professional Summary Dr. Jayant J. Khandare is a globally recognized cancer innovator and entrepreneur with more than two decades of experience at the intersection of chemical engineering, biomaterials, cancer biology, and translational oncology . He is the scientific architect behind multiple breakthrough platforms spanning cancer diagnostics, circulating tumor cell (CTC) technologies, and therapeutic intervention systems , many of which have progressed from concept to clinical and commercial validation. Dr. Khandare has authored over 150+ peer-reviewed scientific publications and is a named inventor on 25+ U.S. patents , reflecting a sustained track record of converting deep science into protectable, scalable intellectual property. His innovations form the foundation of several clinically relevant oncology platforms, including OncoDiscover® (CTC detection and enumeration) and OncoMetastat® (extracorporeal CTC filtration for metastasis mitigation) —technologies designed to address the most critical unmet need in cancer: the prevention and control of metastasis . Internationally trained, Dr. Khandare has held advanced research appointments in the United States and Germany , including postdoctoral work at Wayne State University , Rutgers University , and Freie Universität Berlin , with a strong focus on targeted drug delivery, macromolecular therapeutics, and cancer cell–material interactions . He is an Alexander von Humboldt Foundation Experienced Researcher and a Fellow of the Royal Society of Chemistry (UK) , honors reserved for scientists with sustained international impact. As a scientific founder and operator, Dr. Khandare brings a rare combination of visionary platform thinking and execution discipline . He has successfully built technologies across the full innovation lifecycle—from fundamental materials science and device engineering to translational studies, regulatory pathways, and clinical adoption . Under his leadership, Actorius’ platforms have generated extensive clinical evidence, international publications (ASCO, AACR, ESMO, ISLB), and a growing global IP estate. Dr. Khandare’s entrepreneurial mission is clear and consistent: to create first-in-class technologies that redefine how cancer is detected, monitored, and treated , with a particular focus on systemic disease biology rather than organ-limited paradigms. His work positions Actorius at the forefront of a new category in oncology—interventional cancer diagnostics and therapeutics . Want to be a part of this dynamic team? Visit our Careers section to explore current opportunities that match your interests and expertise. Apply Now

Pune start-up gets US patent for delivering drugs to site-specific organs Press Release 6 March 2023 Pune start-up gets US patent for delivering drugs to site-specific organs The patent was granted to Actorius Innovations and Research and its team that designed capsule shells using natural polymer to obtain a delayed release profile suitable for delivery of drugs to colon and rectum, said Dr Jayant Khandare, founder-director and Chief Scientific Officer of the start-up. Changes in lifestyle and food habits are leading to many colon related diseases including cancers, he said. Delivery of drugs to colorectal site is most challenging as the dosage forms have to prevent the early release of drug in stomach and intestine, he said. This patent (US Patent No. 11596607) is titled "Polymer based formulation for the release of drugs and bioactives at specific GIT sites". Khandare said the technology composition does not involve cumbersome tablet processing, coating and enteric or other polymers. It also reduces processing cost with increased patient compliance, he added. The start-up completed the bio equivalence study which was approved by Drugs Controller General of India (DCGI) in September 2020, Khandare said. Click the link below to read the full article. Know More Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

The deadliest cancer threat may be invisible — just one surviving cell can be enough to restart the disease journey - Dr Jayant Khandare Press Release 24 May 2026 The Hidden Threat of a Single Cell: Dr. Jayant Khandare on How One Circulating Tumor Cell Can Restart the Cancer Journey The deadliest cancer threat may be invisible — just one surviving cell can be enough to restart the disease journey. It takes just one cell to restart the story. In a thought-provoking insight into cancer progression, Dr. Jayant Khandare, Co-founder of Actorius Innovations and a leading researcher in circulating tumor cells, asks a pivotal question: “Can a single cell restart the cancer journey?” 🔗 Read the featured articles: English Article: https://english.dainikjagranmpcg.com/education/6a1285a923ea1/article-19162 Hindi Article: https://www.dainikjagranmpcg.com/national-international/the-hidden-danger-of-a-single-cell-dr-jayant-khandare/article-54101 Explore more research publications: https://www.actorius.in/newsroom #Actorius #DrJayantKhandare #CancerResearch #OncologyInnovation #LiquidBiopsy #OncoDiscover #CancerDiagnostics #HealthcareInnovation #PrecisionMedicine #CTC #CancerAwareness #MedicalResearch #Biotechnology #EarlyDetection #CancerCare Read Article Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

Combined ctDNA and PD-L1 CTC testing improves lung cancer monitoring and response. Publications 3 June 2025 Quadrant of co-occurrence of circulating tumor DNA and PD-L1 expression on circulating tumor cells in monitoring disease aggressiveness and metastasis in lung cancer. Combined ctDNA and PD-L1–positive CTC analysis improves monitoring of metastasis, minimal residual disease, and treatment response in lung cancer. Background Liquid biopsies analyzing circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs) enable minimally invasive monitoring and testing of lung cancer across different stages. Approximately 90% of patients succumb due to metastasis. However, identifying patients with early metastatic signatures remains extremely challenging. In addition, monitoring minimal residual disease (MRD) and identifying patients at risk of recurrence is highly important. While the prognostic role of CTCs in predicting survival has been established in several cancers, the combined role of CTCs and ctDNA in monitoring disease aggressiveness, treatment response, and therapeutic decision-making has not been extensively explored. In this study, we investigated the combined roles of ctDNA and CTCs in monitoring disease aggressiveness and metastasis in lung cancer patients. Methods A cohort of 265 late-stage lung cancer patients was retrospectively analyzed for the co-occurrence of the dual biomarkers ctDNA and CTCs. The results were correlated in a quadrant-based model to assess clinical disease states derived from PET scans and histopathological examination (HPE) findings. Next-generation sequencing (NGS) was performed using the OncoMonitor dual biomarker assay, which includes CTC enumeration with PD-L1 expression analysis. CTC counts were determined using the OncoDiscover Liquid Biopsy Test, approved by CDSCO-India, from 1.5 mL of blood. Results CTC distribution ranged from 1 to 8 cells, with a mean value of 1.22. Among the patients, 75.47% (n = 200) were CTC-positive, and among these, 91.50% (n = 183) exhibited PD-L1 expression on their CTCs, with a mean PD-L1–positive CTC value of 0.99. Both biomarkers were positive (ctDNA⁺/CTC⁺) in 135 patients (50.94%). Only 19 patients (7.17%) were negative for both biomarkers (ctDNA⁻/CTC⁻). Additionally, 43 patients (16.23%) were ctDNA⁺/CTC⁻, while 68 patients (25.66%) were ctDNA⁻/CTC⁺. The ctDNA⁺/CTC⁻ cohort exhibited the highest metastatic rate at 62.8%, followed by the ctDNA⁺/CTC⁺ group at 57.0%. The ctDNA-positive cohort showed the highest proportion of progressive disease (20.2% and 18.6% in CTC⁺ and CTC⁻ subgroups, respectively). Mutations in EGFR, TP53, and KRAS were observed in 62.64% (166/265) of patients. Stable disease was observed in 29.4% of patients when both biomarkers were absent (ctDNA⁻/CTC⁻). Conclusions Overall, the ctDNA-positive cohort demonstrated higher rates of MRD, disease progression, and metastasis, with no cases of stable disease. The combined quadrant analysis of CTC-PD-L1 cells and ctDNA provides a non-invasive approach for monitoring disease progression, treatment response, complete remission, and early metastatic detection in lung cancer patients. View Publication Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe

CK18 variance in CTCs across cancer types necessitates validated enumeration. Publications 9 October 2021 ESMO 2021: Validation of Cytokeratin (CK18) Protein Expression in Epithelial Cell lines and in Circulating Tumor Cells (CTCs) Study shows significant CK18 expression variance across different cancer cell lines and CTCs, highlighting the need for regulated enumeration tools. Background CTCs predict an unfavourable prognosis and outcomes in cancers. Lowering of cytokeratin 18 expression is a hallmark of epithelial mesenchymal transition (EMT). Homogeneity and validation of CK18 expression in cancer cell lines and CTCs originating from distinct solid tumors is indeterminate and may contribute to non-specific counts. We hypothesize that the expression of CK18 in varied cell lines may differ quantitatively, and additionally may exhibit similar trends in CTCs enumerated from different tumor types. Methods CK18 variance in epithelial cell lines (e.g., A549+, MCF-7+, and MEF-) (n=192,269 cells) and CTCs (n=63) of different phenotypes was analyzed and compared. The fluorescence intensity was measured post-immunostaining, using motorized-automated, computer-assisted scanning, and through a customized ImageJ macro tool. The effect of anti-CK18 concentrations (0.06-6 μg/ml) and binding constants (Kb) was measured across all cell lines. CTCs were enumerated from head and neck squamous cell carcinoma (HNSCC) patients' blood samples (CTRI/2018/03/012905) and from clinical samples (e.g., breast, lung, colorectal (CRC), ovarian) using the clinically relevant OncoDiscover platform. Results CK18 mapping revealed diverse fluorescence intensities distribution in three cell lines, as well as in HNSCC, lung, breast, ovarian, and colorectal cancer CTCs (Table). In addition, the protein binding assay showed 8.65 x 10^3 Kb (M^-1) for MCF7 and 7.9 x 10^3 for A549 cells indicating concentration-dependent binding for CK18 expressing proteins on cells and may be varied in CTCs of different cancer types. Compared to the CK- cell line (MEF), the normalized CK18 intensity was higher by 290% and 310%, respectively, in MCF7 (breast) and A549 (lung) cells, demonstrating the variation in CK18 expression. On the other hand, CTCs showed significant diversity in CK18 expression with buccal mucosa revealing the lowest (67%), while CTCs of CRC origin demonstrated the highest expression (320%) (Table). CK18 intensity was represented across the cell lines and on CTCs enumerated from different cancer types. Conclusions Non-regulated CTC enumeration platforms pre-requisite critical validations to eliminate the non-specificity of CTC counts, which are highly imperative to clinical decisions in cancer management. Clinical Trial Identification CTRI/2018/03/012905. Know more Stay One Step Ahead of Cancer. Get the latest news and innovations from Actorius delivered straight to your inbox. Subscribe for regular updates Email* Yes, subscribe me for regular updates. * Subscribe