From:  Targeting tumor transition windows

 Emerging biomarkers for monitoring tumor state transitions during therapy.

Biomarker categoryTechnology/platformBiological information capturedClinical applicationsLimitations
Single-cell transcriptomicsscRNA-seq, high-throughput droplet-based sequencing platformsGene expression states of individual tumor cells; identification of rare adaptive populations such as drug-tolerant persister cells; mapping of cellular state transitions [185, 186]Identification of therapy-resistant subpopulations; characterization of tumor heterogeneity; monitoring transcriptional state transitions during treatmentRequires tumor tissue sampling; high cost; complex computational analysis
Single-cell epigenomicsscATAC-seq, chromatin accessibility profiling, multi-omics single-cell platformsEpigenetic regulatory states, enhancer activity, and transcription factor networks governing tumor cell identity [187, 188]Identification of epigenetic programs driving plasticity and therapy resistance; detection of lineage state transitionsTechnical complexity; limited clinical implementation; dependence on high-quality tissue samples
ctDNALiquid biopsy assays, digital polymerase chain reaction (PCR), next-generation sequencing of plasma DNADetection of tumor-derived mutations and clonal evolution in circulation; dynamic monitoring of tumor genomic alterations during therapy [171, 177]Longitudinal monitoring of treatment response; early detection of emerging resistance mutations; minimal residual disease assessmentLimited ability to capture transcriptional or phenotypic states; sensitivity may vary with tumor burden
CTCsMicrofluidic capture platforms, immunomagnetic enrichment, single-cell analysis of CTCsPhenotypic and molecular characterization of viable tumor cells undergoing dissemination and therapeutic adaptation [189, 190]Monitoring tumor evolution and metastatic potential; assessment of therapy-induced phenotypic transitionsLow abundance in circulation; technical challenges in isolation and characterization
Epigenetic biomarkersDNA methylation profiling, chromatin accessibility assays, enhancer landscape analysisEpigenetic states associated with tumor plasticity, lineage transitions, and drug tolerance [187, 191]Identification of regulatory state changes associated with therapy resistance; potential biomarkers for adaptive tumor statesEpigenetic states may be heterogeneous across tumor regions; clinical standardization still developing
Metabolic biomarkersMetabolomics profiling, metabolic imaging, measurement of metabolic pathway activityMetabolic adaptations occurring during therapeutic stress, including altered nutrient dependencies and mitochondrial function [90, 97]Identification of therapy-induced metabolic vulnerabilities; monitoring tumor metabolic reprogramming during treatmentMetabolic signals can be influenced by systemic physiological factors; interpretation may be complex
Imaging biomarkersfMRI, PET imaging, hypoxia imaging, metabolic imaging tracersSpatial and temporal changes in tumor metabolism, perfusion, hypoxia, and treatment response [187, 192]Non-invasive monitoring of tumor physiology during therapy; detection of dynamic treatment responsesLimited molecular resolution; may not capture cellular-level state transitions

CTCs: circulating tumor cells; ctDNA: circulating tumor deoxyribonucleic acid; fMRI: functional magnetic resonance imaging; PET: positron emission tomography; scATAC-seq: single-cell assay for transposase-accessible chromatin using sequencing; scRNA-seq: single-cell ribonucleic acid sequencing.