TP53 mutations in cancer: Molecular features and therapeutic opportunities (Review).

The tumour suppressor factor p53 plays an essential role in regulating numerous cellular processes, including the cell cycle, DNA repair, apoptosis, autophagy, cell metabolism and immune response. TP53 is the most commonly mutated gene in human cancers. These mutations are primarily non­synonymous changes that produce mutant p53 proteins characterized by loss of function, a dominant negative effect on p53 tetramerisation and gain of function (GOF). GOF mutations not only disrupt the tumour­suppressive activities of p53 but also endow the mutant proteins with new oncogenic properties. Recent studies analysing different pathogenic features of mutant p53 in cancer­derived cell lines have demonstrated that restoring wild­type p53, rather than removing GOF mutations, reduces cancer cell growth. These findings suggest that therapeutic strategies for reactivating wild­type p53 function in cancer cells may bring a greater benefit than approaches halting mutant p53. This approach could involve the use of small molecules, gene therapy and other methods to re­establish wild­type p53 activity. This review describes the complexity of the biological activities of different p53 mutants and summarizes the current therapeutic approaches to restore p53 function.

Wrist fusion with ulnar translocation for aggressive giant cell tumour of distal end radius: Short-term functional and clinico-radiological outcomes.

Background: The distal end radius (DER) is the third most common site of a giant cell tumour (GCT) in bone. GCT is a locally aggressive benign tumour with metastatic potential. The main goals in the management of GCT of DER are the complete removal of the tumours, the prevention of recurrence, and the restoration of the functional wrist. Purpose: This case series reports the clinical and functional outcome of en bloc tumour resection, ulnar translocation and wrist arthrodesis in patients with high-grade GCT of DER. Methods: Sixteen patients with Campanacci grade II & III GCT of distal end radius who underwent the procedure between 2009 and 2018 and had a follow-up of 2 years were included. Patients with distant metastasis, chronic severe systemic illness, operated previously, and follow-up dropouts were excluded from the study. The patient's demographic profile and tumour characteristics were collected at the initial visit. Grip strength, modified MAYO wrist score, and VAS score for wrist pain were collected preoperatively and at 1-year and 2-year follow-ups. Results: The study included predominantly females (13 vs 3). The mean age was 31.5 ± 8 years with a symptom duration of 6.5 ± 3.1 months (range 3-14). The mean resection length was 8.31 ± 1.5 cm (range 6-12). The mean Ulnoradial and ulnocarpal union time was 22.7 ± 8.0 and 17.5 ± 2.3 weeks, respectively. The mean modified Mayo Wrist score was 20.63 ± 9.4, 48.7 ± 5.6 and 60.6 ± 4.0 at preop, 1 year and 2 years follow-up. The mean MSTS score was 22.68 ± 1.8 (range 19-26). Two patients had ulnoradial nonunion. Conclusion: Wrist arthrodesis by ulnar translocation and plate fixation is a viable option in the management of GCT of distal end radius. The short-term clinical and functional outcomes are favourable for restoring adequate wrist function.

Gamma-knife radiosurgery for jugular foramen schwannomas. A systematic review and meta-analysis.

Introduction: Jugular Foramen Schwannomas (JFS) have been traditionally treated with surgical resection with an associated significant post-operative morbidity. Stereotactic radiosurgery has been investigated as potentially minimally invasive alternative to microsurgery. The aim of this study was to provide a systematic review and meta-analysis of the available literature regarding the outcomes of cases of JFS treated with radiosurgery. Methods: A literature review until 28th of March 2023 was performed. All studies looking at the outcomes of radiosurgery for the treatment of JFS were included. Studies including non-vestibular schwannomas without clear distinction of the tumour type were excluded. Risk of bias was assessed using the Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) scale. Results: Eight (8) studies with a total of 375 patients met the inclusion and exclusion criteria and were included in the analysis. Pooled overall tumour control rate was 93.2 % (95 % CI 89.8-96.6) after a weighted mean follow-up of 54.07 months (95 % CI 46.8-61.3). Patient free survival was reported only in 4 studies and ranged from 87 % to 97 % and 76.9-93.8 % in 5 and 10 years respectively. The radiation induced cranial nerve deficits rates after GKRS were 3.6 % (95%CI 1.7, 5.5 %). Conclusion: According to our findings, radiosurgery for JFS has favourable clinical outcomes with a high rate of long-term tumour control and low complication rates.

3D Tumor-Engineered Model Replicating the Osteosarcoma Stem Cell Niche and In Vivo Tumor Complexity.

Osteosarcoma, among all bone sarcomas, remains a challenge despite the unwavering efforts of medical professionals and scientists. To address this, the scientific community is actively pursuing the development of three-dimensional (3D) in vitro models to faithfully replicate the heterogeneity of osteosarcoma, thereby facilitating the reliable preclinical screening of potential therapies. In this study, we present the latest advancements in engineering an in vitro 3D osteosarcoma model comprising enriched Cancer Stem Cells (CSCs) and a hybrid hydroxyapatite-based scaffold (MgHA/CoII). The improvement of the model occurred through two primary steps: (1) serial passaging of sarcospheres as the CSCs enrichment system and (2) the optimization of the structural configuration of the niche in the scaffold. Two injection-mediated approaches of sarcosphere seeding were designed and extensively characterized in vitro and in vivo Chorioallantoic Membrane (CAM) models to explore their biological properties and tumorigenic potential. The combination of the selected enriched-CSCs and custom-made seeding into the scaffold resulted in the development of 3D osteosarcoma models exhibiting tumor-like features in vitro and tumorigenic properties in vivo. The outcomes of this study offer prospects for future endeavors involving more complex systems capable of replicating specific malignant tumor behaviors (metastatic process and drug resistance), pushing the discovery of new therapeutic strategies for clinical applications.

PLXNB1/SEMA4D signals mediate interactions between malignant epithelial and immune cells to promote colorectal cancer liver metastasis.

Distal metastases result from metastatic microenvironment and tumour epithelial cell interactions, the cellular heterogeneity of primary colorectal cancer (CRC) and liver metastases (LM) was evaluated by integrating single-cell sequencing data, and the collected gene expression data from metastatic epithelial cell subsets was used to construct a prognostic model and to identify intercellular receptor-ligand interactions between epithelial and immune cells in CRC and LM. Multiplex immunofluorescence staining, and in vitro wound healing, cell migration and cell apoptosis assays were performed to further explore the biological relevance of identified potential regulatory molecules. In this study, approximately 17 epithelial cell subtypes were detected, with Epi-11 cells being highly expressed in LM tissues compared with CRC samples. Furthermore, patients with high expression of the metastasis-related genetic profile of Epi-11 had a poorer prognosis. By predicting receptor-ligand interactions, Epi-11 cells were found to interact more with myeloid and T/natural killer cells in LM tissues when compared to primary CRC samples, which was mediated by the PLXNB1/SEMA4D axis. In addition, high SEMA4D expression was correlated with decreased overall survival of patients with CRC, whereas PLXNB1 was not. SEMA4D knockdown prevented the migration and promoted the apoptosis of HCT116 cells in vitro. In summary, Epi-11 cells, an important subset of epithelial cells, may drive the LM of CRC and act by crosstalk with immune cells through the PLXNB1/SEMA4D signalling axis.

Evaluation of the Effect of a New Skin Fixation Technique to Avoid Shrinkage of Skin Samples Obtained from Canine Cadavers.

Skin shrinkage begins immediately after surgical incision and is an artefact associated with the excision and fixation of a specimen. Skin shrinkage results in important changes in histologic tissue dimensions and can affect the correct quantification of the histologic tumour-free margin (HTFM). Bilateral and symmetrical circular skin samples with a diameter of 60 mm were taken from the lateral thoracic, flank and femoral regions of dog cadavers, with the samples from one side belonging to the study group and the samples from the same animal from the other side belonging to the control group. The radius and diameter of the specimen were measured immediately after the excision and 10 min later for each sample. The measurements of the study group were taken again after manual re-extension and fixation on a cork plate before formalin fixation and 48 h after formalin fixation. A total of 66 (33 study and 33 control group) samples were collected from 11 canine cadavers. The mean diameter shrinkage after formalin fixation was 18.24% for the control group and 0.64% for the study group. A statistically significant difference between the study and the control group was found (p < 0.001). This method of specimen fixation in the study group avoided skin shrinkage and deformation of the specimen in formalin, which we believe improves the diagnostic accuracy of surgical margins and, thus, reduces the number of false-positive or false-negative HTFM.

Successful surgical management of polyostotic costal fibrous dysplasia-a case report.

Polyostotic costal fibrous dysplasia is a rare cause of chest pain. This report describes a young male with polyostotic fibrous dysplasia with partial vertebral involvement, who presented to us with disabling pain refractory to medical therapy. He was managed successfully with resection of dysplastic ribs, while conserving the asymptomatic vertebral lesion.

Deep Circumflex Iliac Artery Osteoseptocutaneous Flap as a Reconstruction Method for Distal Radius Recurrent Giant Cell Tumour in the Case of a Bilateral Peroneal Magna Artery: An Eight-Year Follow-Up.

A vascularized fibula flap is an option to reconstruct osseous and soft tissue defects involving distal radius malignancy with massive soft tissue involvement. This reconstruction method is a strong anatomical construct for wrist arthrodesis and flexible septocutaneous tissue for closure. However, in rare cases of bilateral peroneal magna artery, a vascularized fibula flap is not a suitable option given its potential risk of limb ischemia. We report the case of a 35-year-old lady with recurrent distal radius giant cell tumor with bilateral peroneal magna artery, whereby a vascularized fibula flap is not a reconstruction option for the distal radius. In this case, we opted to use the deep circumflex iliac artery (DCIA) flap to reconstruct the defect. This case highlights the importance of clinical assessment and Doppler evaluation before harvesting a vascularized fibula graft and the DCIA flap as an alternative option for reconstruction of the distal radius with a good functional outcome eight years post-operation.

Efficacy of immune checkpoint inhibitor combination therapy prior to nephrectomy in advanced renal cell carcinoma: A retrospective pilot study.

Renal cell carcinoma (RCC) affects 10%-20% of patients annually, often with metastases present. This study evaluated the impact of systemic therapy before nephrectomy in patients with unresectable or metastatic renal cell carcinoma (RCC). Patients receiving upfront immune checkpoint inhibitor (ICI) combination therapy showed significantly improved progression-free survival (PFS) compared to nephrectomy alone (2-year PFS: 62.3% vs. 17.4%; p = 0.036), while upfront tyrosine kinase inhibitor (TKI) therapy did not (2-year PFS: 18.2% vs. 12.3%; p = 0.545). Surgery-related outcomes did not differ significantly between groups. ICI therapy maintained tumour reduction rates better than TKI therapy. The study highlights the potential benefits of ICI combination therapy over TKI therapy in advanced RCC, suggesting further research is needed to confirm these findings.

Undetectable pre-radical cystectomy circulating tumour DNA status predicts improved oncological outcomes.

OBJECTIVE: To assess recurrence-free survival (RFS) in patients with undetectable tumour-informed circulating tumour DNA (ctDNA) before radical cystectomy (RC) and evaluate if those who converted from detectable to undetectable ctDNA status after RC have similar RFS outcomes as those with persistently undetectable ctDNA status. PATIENTS AND METHODS: Patients who underwent RC had prospectively and longitudinally collected tumour-informed ctDNA analyses during 2021-2023. ctDNA status was informed from the pre-RC specimen. The minimal residual disease (MRD) window was defined as the initial 90 days after RC. RFS was evaluated using the Kaplan-Meier method. Cox regression analysis was performed to find predictors of disease recurrence. RESULTS: The cohort included 135 patients with 647 ctDNA analyses. The median (interquartile range [IQR]) age was 71 (63-77) years. Over a median (IQR) follow-up of 11 (7-18) months, 41 patients (30%) had a recurrence. Pre-RC undetectable ctDNA status was found in 54 patients (40%). The RFS rates at 6, 12, and 21 months were 98%, 93%, and 82%, respectively. Of 77 patients with undetectable ctDNA status at the MRD window available for conversion dynamics analysis, 43 had persistently undetectable ctDNA status (both at pre-RC and MRD window) and 31 converted from pre-RC detectable to MRD undetectable status (conversion group). The persistently undetectable group had significantly better RFS than the conversion group (log-rank, P < 0.001), with 12-month RFS rates of 97% vs 51%, and 18-month RFS rates of 88% vs 51%, respectively. On Cox multivariate analysis, only the conversion group status predicted disease recurrence. CONCLUSIONS: Patients with undetectable pre-RC ctDNA status have a favourable prognosis and may be candidates for treatment de-escalation. Those with persistently undetectable ctDNA had superior RFS compared to the conversion group. Pre-RC ctDNA status should be incorporated into trials examining ctDNA use in clinical decision-making.

Therapy adherence after interdisciplinary tumour board discussion is associated with improved outcome in soft tissue sarcoma: A Charité Comprehensive Cancer Centre analysis.

Centralising soft tissue sarcoma (STS) treatment in expert centres and implementing comprehensive therapy concepts through interdisciplinary tumour boards (ITB) has led to significant treatment progress. However, our knowledge on the implementation of the ITB recommendations and its impact on patient outcome is limited. In this retrospective analysis, we examined a cohort of 222 adult patients (pts) with primary STS who were presented to the ITB of the Charité Comprehensive Cancer Centre between 2015 and 2020. In localised disease (n = 188), resection was recommended in 71% (n = 134) of pts. The treatment modalities chemotherapy with or without regional deep hyperthermia, and radiotherapy were recommended in 37% (n = 69), 26% (n = 48) and 52% (n = 97), respectively. Complex multidisciplinary concepts were established in 29% (n = 54) including ≥3 treatment modalities. Only partial adherence, either by choice of patient or treating physician, was associated with a higher risk of both progression (HR 4.0 95%-CI 1.6-9.7 p < .01) and mortality (HR 5.3 95%-CI 1.7-16.4 p < .01). Pts inable to follow the ITB recommendations due to complications or rapid progression showed a high-risk profile with increased mortality and progression rates (HR 18.1 95%-CI 8.5-38.2 p < .001; HR 21.5 95%-CI 8.5-54.7 p < .001). To our knowledge, this represents the first German Comprehensive Cancer Centre analysis of therapy adherence in STS. It provides further real-world evidence that full adherence to ITB recommendations and the ability to adhere to them are of prognostic value for patient outcome and underlines the importance of interdisciplinary decision-making and treatment planning for STS patients.

DNA methylation-array interlaboratory comparison trial demonstrates highly reproducible paediatric CNS tumour classification across 13 international centres.

AIMS: DNA methylation profiling, recently endorsed by the World Health Organisation (WHO) as a pivotal diagnostic tool for brain tumours, most commonly relies on bead arrays. Despite its widespread use, limited data exist on the technical reproducibility and potential cross-institutional differences. The LOGGIC Core BioClinical Data Bank registry conducted a prospective laboratory comparison trial with 12 international laboratories to enhance diagnostic accuracy for paediatric low-grade gliomas, focusing on technical aspects of DNA methylation data generation and profile interpretation under clinical real-time conditions. METHODS: Four representative low-grade gliomas of distinct histologies were centrally selected, and DNA extraction was performed. Participating laboratories received a DNA aliquot and performed the DNA methylation-based classification and result interpretation without knowledge of tumour histology. Additionally, participants were required to interpret the copy number profile derived from DNA methylation data and conduct DNA sequencing of the BRAF hotspot p.V600 due to its relevance for low-grade gliomas. Results had to be returned within 30 days. RESULTS: High technical reproducibility was observed, with a median pairwise correlation of 0.99 (range 0.94-0.99) between coordinating laboratory and participants. DNA methylation-based tumour classification and copy number profile interpretation were consistent across all centres, and BRAF mutation status was accurately reported for all cases. Eleven out of 12 centres successfully reported their analysis within the 30-day timeframe. CONCLUSION: Our study demonstrates remarkable concordance in DNA methylation profiling and profile interpretation across 12 international centres. These findings underscore the potential contribution of DNA methylation analysis to the harmonisation of brain tumour diagnostics.

A review of acute symptomatic seizures during awake craniotomy for tumour resection.

PURPOSE: Awake craniotomy (AC) is a procedure often performed concomitantly with direct electrical cortical stimulation (DES) and electrocorticography (ECoG) during functional brain mapping. Patients undergoing AC are at risk of acute symptomatic seizures, including intraoperative (IS) and early postoperative seizures (EPS) which can lead to higher risk of morbidity. Predicting those who are at risk of IS and EPS could alert clinicians and provide the ability to closely monitor and consider management changes in the acute setting to prevent seizures. MATERIALS AND METHODS: This is a narrative review of previous studies on IS and EPS during awake craniotomy, including a summary of studies from our center using a novel circular grid electrode. RESULTS AND CONCLUSIONS: There are a number of clinical features with variable association with a higher risk of EPS and IS. Surgeries involving the anterior and central head regions are a risk factor for IS. EPS is more likely to occur in patients with perioperative intracranial hemorrhage. Improving grid/electrode technology for ECoG can allow for better sensitivity of detecting epileptiform activity which can help to diagnose and predict perioperative seizures.

Targeting Bone Tumours with 45S5 Bioactive Glass.

Despite advances in treatment modalities, bone tumour therapies still face significant challenges. Severe side effects of conventional approaches, such as chemo- and radiation therapy, result in poor survival rates and high tumour recurrence rates, which are the most common issues that need to be improved upon. The aim of this study was to evaluate the therapeutic properties of 45S5 bioactive glass (BG) for targeting bone tumours. The viability of the cells derived from osteosarcoma, chondrosarcoma, and giant cell tumours was significantly reduced in the presence of 45S5-BG. In contrast, the viability of non-malignant osteoblast-like cells, chondrocytes, and bone marrow-derived stromal cells was not or only slightly affected. While alterations to the particle surface induced by heat treatment, acid etching, or incubation in a simulated body fluid had only minor effects on cytotoxicity, reducing the particle size or sintering the material significantly improved the cytotoxic effect of 45S5-BG. Further, using a chicken chorioallantoic membrane assay, the co-transplantation of 45S5-BG resulted in a significant reduction in tumour formation in vivo. Given the known positive effects of BGs on bone regeneration, our findings suggest that 45S5-BG holds great potential for the development of new and effective bone tumour therapies, with minimal side effects on non-malignant cells and simultaneous contribution to bone healing.

PD-L1+ Lymphocytes Are Associated with CD4+, Foxp3+CD4+, IL17+CD4+ T Cells and Subtypes of Macrophages in Resected Early-Stage Non-Small Cell Lung Cancer.

The non-canonical PD-L1 pathway revealed that programmed-death ligand 1 (PD-L1) expression in immune cells also plays a crucial role in immune response. Moreover, immune cell distribution in a tumour microenvironment (TME) is pivotal for tumour genesis. However, the results remain controversial and further research is needed. Distribution of PD-L1-positive (PD-L1+) tumour-infiltrating lymphocytes in the context of TME was assessed in 72 archival I-III stage surgically resected NSCLC tumour specimens. Predominant PD-L1+ lymphocyte distribution in the tumour stroma, compared to islets, was found (p = 0.01). Higher PD-L1+ lymphocyte infiltration was detected in smokers due to their predominance in the stroma. High PD-L1+ lymphocyte infiltration in tumour stroma was more common in tumours with higher CD4+ T cell infiltration in islets and stroma, Foxp3+CD4+ T cell infiltration in islets and lover M1 macrophage infiltration in the stroma (p = 0.034, p = 0.034, p = 0.005 and p = 0.034 respectively). Meanwhile, high PD-L1+ lymphocyte infiltration in islets was predominantly found in tumours with high levels of IL-17A+CD4+ T cells in islets and Foxp3+CD4+ T cells in islets and stroma (p = 0.032, p = 0.009 and p = 0.034, respectively). Significant correlations between PD-L1+ lymphocytes and tumour-infiltrating CD4+, Foxp3+CD4+, IL-17A+CD4+ T cells and M2 macrophages were found. An analysis of the tumour-immune phenotype revealed a significant association between PD-L1 expression and IL17+CD4+ and Foxp3+CD4+ immune phenotypes. PD-L1+ lymphocytes are associated with the distribution of CD4+, Foxp3+CD4+, IL17A+CD4+ T cells, M1 and M2 macrophages in TME of resected NSCLC.

Analysis of Fibropapillomatosis in Roe Deer (Capreolus capreolus) Confirms High Content of Heavy Metals.

In recent decades, there has been an increase in European wild ungulate populations, often associated with a decline in health and spread of disease. This is true for the roe deer (Capreolus capreolus), the most common European cervid, with populations apparently affected by fibropapillomatosis, an increasingly common cancer. To date, however, there has been little research into this disease, thus many interactions remain unclear and descriptions of tumour composition are poorly validated. The main aim of the present study was to evaluate the presence and concentration of toxic heavy metals in roe deer skin tumours. Our results confirmed the presence of virtually all the metals tested for, i.e., Pb, Hg, Cd, As, Cr, Mn, Al, Co, Cu, Ni, Se, Zn, and Fe, with the highest average concentrations found for Cr (0.99 mg/kg-1 ± 2.23 SD), Cd (0.03 mg/kg-1 ± 0.03 SD), and Hg (0.02 mg/kg-1 ± 0.02 SD), exceeding FAO limits for meat from slaughtered animals. We also observed a significant positive relationship between heavy metal concentration and age, especially for Pb, As, Hg, Mn, Se, Al, Zn, and Ni. Our findings provide a strong baseline for further research on the impact of fibropapillomatosis, not only on the welfare and health status of game but also on the final consumer of venison, which in many respects is regarded as a high-quality, ecological, and renewable wild resource. While deer with this disease are not considered qualitatively or medically defective, they could represent a potential reservoir of substances toxic to humans and could affect substance levels in adjacent tissues or the animal as a whole.

The Multimodality Management of Malignant Peripheral Nerve Sheath Tumours.

Malignant peripheral nerve sheath tumours (MPNST) are aggressive sarcomas that have nerve sheath differentiation and can present at any anatomical site. They can arise from precursor neurofibroma in the context of neurofibromatosis type 1 (NF1) or as de novo and sporadic tumours in the absence of an underlying genetic predisposition. The primary therapeutic approach is most often radical surgery, with non-surgical modalities playing an important role, especially in locally advanced or metastatic cases. The aim of multimodality approaches is to optimize both local and systemic control while keeping to a minimum acute and late treatment morbidity. Advances in the understanding of the underlying biology of MPNSTs in both sporadic and NF-1-related contexts are essential for the management and implementation of novel therapeutic approaches.

Evolution of Liquid Biopsies for Detecting Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy characterised by late diagnosis and poor prognosis. Despite advancements, current diagnostic and prognostic strategies remain limited. Liquid biopsy techniques, including circulating tumour DNA (ctDNA), circulating tumour cells (CTCs), circulating tumour exosomes, and proteomics, offer potential solutions to improve PDAC diagnosis, prognostication, and management. A systematic search of Ovid MEDLINE identified studies published between 2019 and 2024, focusing on liquid biopsy biomarkers for PDAC. A total of 49 articles were included. ctDNA research shows some promise in diagnosing and prognosticating PDAC, especially through detecting mutant KRAS in minimal residual disease assays. CTC analyses had low sensitivity for early-stage PDAC and inconsistent prognostic results across subpopulations. Exosomal studies revealed diverse biomarkers with some diagnostic and prognostic potential. Proteomics, although relatively novel, has demonstrated superior accuracy in PDAC diagnosis, including early detection, and notable prognostic capacity. Proteomics combined with CA19-9 analysis has shown the most promising results to date. An update on multi-cancer early detection testing, given its significance for population screening, is also briefly discussed. Liquid biopsy techniques offer promising avenues for improving PDAC diagnosis, prognostication, and management. In particular, proteomics shows considerable potential, yet further research is needed to validate existing findings and comprehensively explore the proteome using an unbiased approach.

The Role of Liquid Biopsy in Gastroenteropancreatic Neuroendocrine Neoplasms.

Neuroendocrine neoplasms incidence has been increasing, arising the need for precise and early diagnostic tools. Liquid biopsy (LB) offers a less invasive alternative to tissue biopsy, providing real-time molecular information from circulating tumour components in body fluids. The aim of this review is to analyse the current evidence concerning LB in NENs and its role in clinical practice. We conducted a systematic review in July 2024 focusing on LB applications in NENs, including circulating tumour cells (CTCs), circulating tumour DNA (ctDNA), micro RNA (miRNA), messenger RNA (mRNA) and extracellular vesicles. Sixty-five relevant articles were analysed. The LB showed potential in diagnosing and monitoring NENs. While CTCs face limitations due to low shedding, ctDNA provides valuable information on high-grade neoplasms. MiRNA and mRNA (e.g., the NETest) offer high sensitivity and specificity for diagnosis and prognosis, outperforming traditional markers like chromogranin A. The LB has significant potential for NEN diagnosis and monitoring but lacks widespread clinical integration due to limited prospective studies and guidelines, requiring further validation. Advances in sequencing technologies may enhance the clinical utility of LB in NENs. Future research should focus on refining LB methods, standardising protocols and exploring applications in high-grade NENs.

Single-Cell Analysis of Bone-Marrow-Disseminated Tumour Cells.

Metastasis frequently targets bones, where cancer cells from the primary tumour migrate to the bone marrow, initiating new tumour growth. Not only is bone the most common site for metastasis, but it also often marks the first site of metastatic recurrence. Despite causing over 90% of cancer-related deaths, effective treatments for bone metastasis are lacking, with current approaches mainly focusing on palliative care. Circulating tumour cells (CTCs) are pivotal in metastasis, originating from primary tumours and circulating in the bloodstream. They facilitate metastasis through molecular interactions with the bone marrow environment, involving direct cell-to-cell contacts and signalling molecules. CTCs infiltrate the bone marrow, transforming into disseminated tumour cells (DTCs). While some DTCs remain dormant, others become activated, leading to metastatic growth. The presence of DTCs in the bone marrow strongly correlates with future bone and visceral metastases. Research on CTCs in peripheral blood has shed light on their release mechanisms, yet investigations into bone marrow DTCs have been limited. Challenges include the invasiveness of bone marrow aspiration and the rarity of DTCs, complicating their isolation. However, advancements in single-cell analysis have facilitated insights into these elusive cells. This review will summarize recent advancements in understanding bone marrow DTCs using single-cell analysis techniques.