Non-Hodgkin lymphoma (NHL) encompasses a diverse group of hematologic malignancies originating from lymphoid tissues. It is characterized by the malignant proliferation of B-cells, T-cells, or natural killer (NK) cells. NHL can be broadly categorized into indolent (slow-growing) and aggressive (fast-growing) types, with a range of clinical presentations and prognoses. The heterogeneity of NHL makes diagnosis and treatment challenging, necessitating advanced diagnostic and molecular techniques. Globally, NHL is a significant health concern with varying incidence and mortality rates by region and demographics. Recent American Cancer Society statistics rank NHL as the ۷th most common cancer in the U.S., with about ۸۲,۰۰۰ new cases and ۲۱,۰۰۰ deaths expected in ۲۰۲۴. The incidence rate is approximately ۲۰ per ۱۰۰,۰۰۰ people per year, showing an increasing trend over recent decades. The mortality rate associated with NHL also highlights the severity of the disease, with a five-year survival rate of around ۷۲% overall. However, this rate varies significantly depending on the NHL subtype, stage at diagnosis, and response to treatment. Aggressive forms of NHL generally have a poorer prognosis compared to indolent types, underscoring the need for early and accurate diagnosis to improve outcomes. In light of these challenges, advanced diagnostic techniques such as exosome analysis, miRNA profiling, and
next-generation sequencing (NGS) are becoming increasingly critical in enhancing the precision of NHL diagnosis and tailoring individualized treatment plans. The integration of exosome analysis, miRNA profiling, and NGS has revolutionized NHL diagnostics, enhancing disease understanding and personalized treatment. Exosomes, nano-sized vesicles from lymphoma cells, offer non-invasive tumor biomarker capture, carrying miRNAs, proteins, and nucleic acids reflecting tumor changes. Advantages include liquid biopsy applications, reducing invasive procedures and providing real-time tumor insights. Challenges include variability in exosome isolation techniques, standardization difficulties across labs, and biological heterogeneity complicating data interpretation. MiRNA profiling leverages the stability and regulatory roles of miRNAs to identify dysregulated expression patterns associated with NHL. Advantages of this technique include high specificity for disease-associated biomarkers and the potential for identifying miRNAs that can predict disease progression and response to therapy. However, limitations include variability in miRNA expression across different studies and patient populations and the challenge of distinguishing between miRNA profiles of NHL and those of other lymphoid disorders. Furthermore, the lack of standardized methodologies for miRNA quantification can impact reproducibility and clinical utility. NGS offers a comprehensive view of NHL’s genomic landscape by detecting various genetic alterations, including mutations, insertions, deletions, and copy number variations. Its advantages include identifying novel genetic drivers and therapeutic targets, guiding personalized treatments, and detecting multiple alterations simultaneously. However, NGS is limited by high costs, extensive computational needs, and the requirement for advanced bioinformatics tools. Additionally, interpreting the clinical significance of numerous genetic variants, especially distinguishing between pathogenic mutations and benign polymorphisms, remains challenging. Collectively, while the combination of exosome analysis, miRNA profiling, and NGS represents a promising frontier in NHL diagnosis, each method has its advantages and limitations. Addressing these challenges through technological advancements and standardization efforts is crucial for optimizing diagnostic accuracy and improving patient outcomes.