Circulating tumor DNA‐based liquid biopsy is increasingly integrated into cancer care, particularly for detecting minimal residual disease, yet patient perspectives remain poorly understood. This single‐center, cross‐sectional, mixed‐methods study assessed recall, knowledge, and attitudes toward liquid biopsy and circulating tumor DNA among 195 patients (median age 65 years; 63% female) undergoing cancer treatment or follow‐up, supplemented by focus groups. Although 65% reported prior liquid biopsy, this was confirmed in only 44%, and only 10% had heard of circulating tumor DNA (ctDNA). Correct recall and knowledge on ctDNA were higher among patients with neuroendocrine neoplasms and those with postgraduate education. In hypothetical scenarios assuming proven clinical validity and utility, 62–80% supported liquid biopsy‐guided clinical action, 74% were comfortable with liquid biopsy replacing tissue biopsy, and 79% accepted its use to guide adjuvant treatment. Most patients anticipated reassurance rather than stress, and 85% expected improved quality of life with home‐ or general practitioner‐based blood sampling. Overall, patients showed high acceptance of clinically actionable liquid biopsy use, driven by trust and perceived clinical benefit.
Impaired protein folding in the endoplasmic reticulum (ER) causes ER stress and activates the unfolded protein response (UPR), which can induce apoptosis during prolonged ER stress. ER stress is therapeutically targeted in plasma cell malignancies, such as multiple myeloma (MM), but these treatments remain noncurative, and UPR signaling in B‐cell malignancies is incompletely understood. A targeted CRISPR‐Cas9 screen in RPMI‐8226 cells exposed to thapsigargin identified known ER stress regulators (XBP1, IRE1, and calreticulin) and the translation factor eIF5A (eukaryotic translation initiation factor 5A), whose role in ER stress remains unclear. Competition assays showed that eIF5A depletion is detrimental under basal conditions but confers a proliferation advantage during thapsigargin treatment. Inducible EIF5A knockouts confirmed reduced sensitivity specifically to thapsigargin, loss of eIF5A had no or sensitizing effects on cell death induced by other ER stress drugs. Polysome profiling revealed that not general translation, but the translation of specific mRNA transcripts coding for UPR components and apoptotic proteins was affected by eIF5A depletion. Correspondingly, EIF5A KO cells showed reduced thapsigargin‐induced UPR activation. In conclusion, these findings indicate that loss of eIF5A alters the stress response in MM cells.
Signals emanating from the B cell receptor (BCR) are vital for driving cell survival and proliferation in chronic lymphocytic leukaemia (CLL). BCR‐mediated signals in CLL cells promote nuclear exclusion and inactivation of the transcription factor forkhead box protein O1 (FOXO1) and transient upregulation of FOXO1 expression. To investigate the mechanisms that regulate FOXO1 inactivation, we focused on the role of deubiquitinase (DUB) family proteins. Stratifying expression of individual DUB proteins, we established that ubiquitin‐specific protease 7 (USP7), USP8, USP9x, USP10 and USP14 are significantly upregulated compared to healthy B cells. USP7 interacts with FOXO1 in primary CLL cells, suggesting that modulation of USP7 activity may impact FOXO1 function. Using selective USP7 inhibitors, P5091 and HBX19818, or USP7 knockdown, combined with ibrutinib, we demonstrated enhanced FOXO1 nuclear localisation over ibrutinib alone and increased FOXO1 activity, leading to modulation of selected FOXO target genes. This resulted in reduced CLL cell viability and enhanced cell cycle arrest in MEC1 cells, largely reversed by FOXO1 knockdown. These findings suggest that dual targeting of BCR signalling and USP7 activity can harness FOXO1‐mediated anti‐tumour activity and may represent a novel therapeutic approach for CLL treatment.
Angiogenesis is a major driver of tumor progression and a validated therapeutic target in highly vascularized cancers such as clear cell renal cell carcinoma. However, the clinical benefit of antivascular endothelial growth factor therapies is limited by the emergence of resistance. We identified a vascular endothelial growth factor A splice variant, termed Lymphatic and Vascular Resistance Factor (LVRF), which promotes both angiogenesis and lymphangiogenesis and is upregulated following anti‐angiogenic treatment. To therapeutically target this pathway, we generated fully human monoclonal antibodies against LVRF. Lead antibodies displayed high specificity and nanomolar affinity and effectively inhibited LVRF‐driven biological activities. Anti‐LVRF antibodies reduced renal carcinoma cell invasion, inhibited vascular and lymphatic endothelial responses, and suppressed lymphatic endothelial cell sprouting. In vivo , treatment significantly reduced the growth of bevacizumab‐resistant renal carcinoma xenografts, induced regression of blood and lymphatic vessels, and decreased metastatic dissemination in zebrafish models. These findings identify LVRF as a mediator of resistance to anti‐angiogenic therapy and support therapeutic targeting of LVRF as a novel strategy for vascularized tumors resistant to current antivascular endothelial growth factor treatments.
Transcription factors (TFs) are often considered undruggable due to their structural complexity. mRNA technologies can overcome this therapeutic limitation by enabling dynamic protein replacement therapies. Here, we explore the TF activity of LCOR (Ligand‐dependent corepressor), which suppresses tumor growth, at least in part by inducing the antigen presentation machinery (APM) in tumor cells and constraining cellular plasticity. These effects facilitate tumor immune recognition and immune‐mediated tumor eradication. We used poly β‐(amino esters) (pBAE) nanoparticles (NPs) for local delivery of Lcor mRNA in multiple breast cancer models. pBAE‐NPs efficiently encapsulate mRNA and facilitate cellular uptake, achieving optimal endosomal escape, resulting in high transfection efficiency in vitro and in vivo , restoring LCOR function in tumor cells and engaging their APM. In preclinical models, intratumoral delivery of Lcor mRNA reduced tumor growth. Importantly, the combination of Lcor mRNA‐loaded RH‐pBAE‐NPs with anti‐PDL1 or anti‐CTLA4 immunotherapies eradicated most of the tumors in our preclinical TNBC model. Overall, our nanotherapeutic strategy emerges as an innovative TF‐replacement therapy in a lock‐in/wipe‐out fashion, leveraging the immunogenic effects of LCOR to eradicate breast cancer tumors when combined with immunotherapy.
