tbuckley/cabot-search huggingface dataset:
Genetically-modified, redirected T cells target hepatitis B surface antigen-positive hepatocytes and hepatocellular carcinoma lesions in a clinical setting
Authors: Xueshuai Wan, Karin Wisskirchen, Tao Jin, Lu Yang, Xiaorui Wang, Xiang’an Wu, Fang Liu, Yu Wu, Christy Ma, Yong Pang, Qi Li, Ke Zhang, Ulrike Protzer, Shunda Du
Journal: Clinical and Molecular Hepatology (2024)

Hepatitis B virus (HBV)-DNA integration in HBV-related hepatocellular carcinoma (HBV-HCC) can be targeted by HBV-specific T cells. SCG101 is an autologous, HBV-specific T-cell product expressing a T-cell receptor (TCR) after lentiviral transduction recognizing the envelope-derived peptide (S20-28) on HLA-A2. We here validated its safety and efficacy preclinically and applied it to an HBV-related HCC patient (NCT05339321). Good Manufacturing Practice-grade manufactured cells were assessed for off-target reactivity and functionality against hepatoma cells. Subsequently, a patient with advanced HBV-HCC (Child-Pugh class A, Barcelona Clinic Liver Cancer stage B, Eastern Cooperative Oncology Group performance status 0, hepatitis B e antigen-, serum hepatitis B surface antigen [HBsAg]+, HBsAg+ hepatocytes 10%) received 7.9×107 cells/kg after lymphodepletion. Safety, T-cell persistence, and antiviral and antitumor efficacy were evaluated. SCG101, produced at high numbers in a closed-bag system, showed HBV-specific functionality against HBV-HCC cells in vitro and in vivo. Clinically, treatment was well tolerated, and all adverse events, including transient hepatic damage, were reversible. On day 3, ALT levels increased to 1,404 U/L, and concurrently, serum HBsAg started decreasing by 3.84 log10 and remained <1 IU/mL for over six months. HBsAg-expressing hepatocytes in liver biopsies were undetectable after 73 days. The patient achieved a partial response according to modified RECIST with a >70% reduction in target lesion size. Transferred T cells expanded, developed a stem cell-like memory phenotype, and were still detectable after six months in the patient's blood. SCG101 T-cell therapy showed encouraging efficacy and safety in preclinical models and in a patient with primary HBV-HCC and concomitant chronic hepatitis B with the capability to eliminate HBsAg+ cells and achieve sustained tumor control after single dosing.
Mitochondrial quality control in human health and disease
Authors: Bo-Hao Liu, Chen-Zhen Xu, Yi Liu, Zi-Long Lu, Ting-Lv Fu, Guo-Rui Li, Yu Deng, Guo-Qing Luo, Song Ding, Ning Li, Qing Geng
Journal: Military Medical Research (2024)

Abstract Mitochondria, the most crucial energy-generating organelles in eukaryotic cells, play a pivotal role in regulating energy metabolism. However, their significance extends beyond this, as they are also indispensable in vital life processes such as cell proliferation, differentiation, immune responses, and redox balance. In response to various physiological signals or external stimuli, a sophisticated mitochondrial quality control (MQC) mechanism has evolved, encompassing key processes like mitochondrial biogenesis, mitochondrial dynamics, and mitophagy, which have garnered increasing attention from researchers to unveil their specific molecular mechanisms. In this review, we present a comprehensive summary of the primary mechanisms and functions of key regulators involved in major components of MQC. Furthermore, the critical physiological functions regulated by MQC and its diverse roles in the progression of various systemic diseases have been described in detail. We also discuss agonists or antagonists targeting MQC, aiming to explore potential therapeutic and research prospects by enhancing MQC to stabilize mitochondrial function.
Inhibition of NOTCH4 sensitizes FLT3/ITD acute myeloid leukemia cells to FLT3 tyrosine kinase inhibition
Authors: Ruiqi Zhu, Courtney M. Shirley, S. Haihua Chu, Li Li, Bao H. Nguyen, Jaesung Seo, Min Wu, Tessa Seale, Amy S. Duffield, Louis M. Staudt, Mark Levis, Yu Hu, Donald Small
Journal: Leukemia (2024)
Re: Srikala S. Sridhar, Thomas Powles, Miguel. Á. Climent Durán, et al. Avelumab First-line Maintenance for Advanced Urothelial Carcinoma: Analysis from JAVELIN Bladder 100 by Duration of First-line Chemotherapy and Interval Before Maintenance. Eur Urol 2024:85:154–63
Authors: Weimin Zhou, Xiongbing Zu, Jiao Hu
Journal: European Urology (2024)
Challenging the Urologist of the Future: Time for a Change?
Authors: Fabian P. Stangl, Isabel Heidegger, Pia Kraft, Rianne J.M. Lammers, Mark Horvath, Muslim Dogan Deger, Elvira Polo-Alonso, Mike Wenzel, Andres Affentranger, Katrine Schou-Jensen, Lobna Ali, Robert A. Keenan, Anna Cadenar, Laura Gallardo-Zamora, Josias Franco Alva, Domenico Trerè, Georgios Tsampoukas, Peter Hoyerup, Clara Cerrato, Nicolas Raison, Sabine Brookman-May, Juan Luis Vásquez
Journal: European Urology (2024)
Adjuvant Everolimus in Patients with Completely Resected, Very High-risk Renal Cell Carcinoma of Clear Cell Histology: Results from the Phase 3 Placebo-controlled SWOG S0931 (EVEREST) Trial
Authors: Primo N. Lara, Catherine Tangen, Elisabeth I. Heath, Shuchi Gulati, Mark N. Stein, Maxwell Meng, Ajjai Shivaram Alva, Sumanta K. Pal, Igor Puzanov, Joseph I. Clark, Toni K. Choueiri, Neeraj Agarwal, Robert Uzzo, Naomi B. Haas, Timothy W. Synold, Melissa Plets, Ulka N. Vaishampayan, Brian M. Shuch, Seth Lerner, Ian M. Thompson, Christopher W. Ryan
Journal: European Urology (2024)
Ketone-based metabolism and signalling in heart failure with preserved ejection fraction (HFpEF)
Authors: R Wang, F Capone, F Capone, L Luo, L L, D Keller, D Keller, S Jung, S Jung, F Pauline, F Pauline, P Mertins, P Mertins, C Sotomayor-Flores, C Sotomayor-Flores, D Bode, D Bode, N Grunert, N Grunert, G Schiattarella, G Schiattarella
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): DZHK Deutsche Forschungsgemeinschaft (DFG-SFB_1470_A02) Cardiac metabolic remodelling in heart failure with preserved ejection fraction (HFpEF) underlies the pathogenesis of this syndrome. Yet, our understanding of metabolic alterations occurring in HFpEF hearts is still limited. Ketone bodies (KBs) – β-hydroxybutyrate (β-OHB) in particular – can become a major, alternative source of fuel for failing hearts. However, the biological relevance of KBs goes beyond their role as energy carrier. We used a ‘two-hit’ mouse model of HFpEF induced by high-fat-diet feeding and L-Name, aiming to evaluate metabolic alteration among various energy substrates (e.g., glucose, fatty acids), in particular the role of β-OHB in HFpEF hearts. Here, we showed that HFpEF heart exhibited impaired glucose oxidation and increased fatty acid oxidation. Impaired glucose oxidation was attributed to the reduced pyruvate dehydrogenase (PDH) activity and increased phospho-PDH expression via the activation of PDH kinase 4 (PDK4). The oxidation of β-OHB was reduced as a result of decreased β-OHB dehydrogenase (BDH1) and succinyl-coA-3-oxaloacid CoA transferase (SCOT) expression. Furthermore, we showed that β-OHB governs mitochondrial function and myocardial bioenergetics in HFpEF, acting as protein modifiers through β-hydroxybutyrylation (Kbhb) – a lysine-based post-translational modification (PTM). Immunoblot analysis for pan- β-hydroxybutyryllysine revealed a significant increase in Kbhb levels in the heart of ‘two-hit’ HFpEF mice compared to control mice. To identify the specific proteins and sites of Kbhb in the heart, Kbhb peptides from control and HFpEF mouse hearts were isolated by immunoaffinity enrichment using a pan-Kbhb antibody-conjugated beads. Volcano plot and gene ontology analysis showed that the most Kbhb-modified proteins in HFpEF hearts are mitochondrial enzymes. Glutamate oxaloacetate transaminase (GOT2) is one of the top hits, participating in the TCA cycle and malate-aspartate shuttle (MAS) in the mitochondria. We further demonstrated that GOT2 activity and MAS were enhacned in the mitochondrial of HFpEF hearts. To conclude, in the heart of ‘two-hit’ model of HFpEF, the oxidation of pyruvate was impaired resulting from decreased PDH activity. The oxidation of β-OHB was also reduced due to the down-regulated BDH1 activity. Instead of oxidation, Kbhb was highlighted as the primary mode of KB utilisation in HFpEF, affecting the GOT2 enzyme activity in MAS pathways. Our results shed light on identifying a novel role of KBs, which can facilitate the understanding of pathogenesis in HFpEF.Ketone-based metabolism and signalling
Nuclear eNOS interacts with and S-nitrosates ADAR1 to modulate endothelial gene expression
Authors: X Zhou, S Guenther, K Kuenne, I Wittig, I Fleming, M Siragusa
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Collaborative Research Centre (CRC 1366); Chinese Scholarship Council (CSC) Introduction and Aim Nitric oxide (NO) generated by the endothelial NO synthase (eNOS) regulates vascular tone and endothelial homeostasis to counteract vascular inflammation. The effects of NO are attributable to its interaction with heme-containing proteins e.g., soluble guanylyl cyclase, or to the S-nitrosation of proteins. Most eNOS is localized to the cell membrane or the Golgi apparatus, however, it has also been detected in the nucleus. In this study, we assessed the role of nuclear eNOS in endothelial cells. Methods and Results Confocal microscopy studies confirmed the presence of eNOS in the nucleus of unstimulated human and murine endothelial cells (ex vivo and in vitro) and stimulation with vascular endothelial growth factor (VEGF) increased eNOS nuclear translocation. Co-immunoprecipitation studies coupled with proteomics revealed the VEGF-dependent association of nuclear eNOS with 81 proteins involved in RNA binding and processing. Among these, nuclear eNOS bound double-stranded RNA-specific adenosine deaminase 1 (ADAR1), an enzyme involved in RNA editing via the deamination of adenosine (A) to inosine (I) in double-stranded RNA (dsRNA). ADAR1 was S-nitrosated in human endothelial cells and the knockdown of eNOS was associated with an increase in dsRNA (immunofluorescence) and altered ADAR1-mediated A-I editing (RNA seq). The accumulation of dsRNA in human endothelial cells lacking eNOS led to the activation of the interferon α/β signalling pathway (e.g., MX2, OASL, OAS1/2/3, RNASEL) and the downregulation of cell cycle-related genes. As a result, growth factor-stimulated cell proliferation was abrogated and basal as well as tumour necrosis factor- or H2O2-induced cell death were increased. Furthermore, the increased dsRNA in endothelial cells lacking eNOS elicited the activation of interferon-induced, dsRNA-activated protein kinase, which in turn phosphorylated the eukaryotic translation initiation factor 2A. Accordingly, new protein synthesis was inhibited following eNOS knockdown (immunofluorescence and mass spectrometry). Conclusion These results demonstrated that eNOS/NO signalling regulates endothelial type I interferon signalling and affects translation, cell cycle and cell death through S-nitrosation of ADAR1.
Human in vitro disease modeling of ATTR cardiac amyloidosis
Authors: J Qin, I Dokter, Y Qiang, Z Lei, J Sluijter, M Oerlemans, A Van Mil
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Private company. Main funding source(s): Pfizer Alnylam Transthyretin amyloid cardiomyopathy (ATTR-CM) is a progressive disease with deposition of TTR amyloid fibrils in the myocardium and cardiac dysfunction with dismal prognosis. In ATTR-CM, either genetic mutations (variant; v) or ageing (wild-type; wt) leads to TTR instability. Late diagnosis and a lack of human disease models limit disease understanding and therapeutic discovery. This study aimed to model ATTR-CM in vitro by exposing various cardiac cell types and multicellular 3D cardiac spheroids to TTR fibrils, characterizing disease phenotypes and providing insights into the cellular mechanisms of disease progression. Human recombinant TTR (WT, V30M, V122I) and acid induced fibrils were generated and characterized. Seeding hiPSC-cardiomyocytes (CMs) and endothelial cells (ECs) on TTR fibrils resulted in cell viability reduction. Confocal microscopy revealed the extracellular attachment of TTR fibrils, leading to sarcomere disorganization, prolonged calcium transients and disrupted electromechanical coupling in hiPSC-CMs, and declined migration capacity of ECs showing aberrant cell morphology. Similarly, cardiac spheroids incorporating TTR fibrils resulted in decreased cell viability and prolonged calcium transients with lower amplitude. Our models show the discovery of early cellular ATTR-CM phenotypes, providing a novel approach to study the disease progression, potentially facilitating the identification of novel therapeutic targets and biomarkers.Cytotoxicity of TTR fibrils on hiPS-CMs
The novel bispecific anti-NLRP3 inflammasome antibody InflamAb inhibits atherosclerosis in apolipoprotein E-deficient mice
Authors: L Delfos, M A C Depuydt, M N A Bernabe Kleijn, A C Foks, J Kuiper, M Chemaly, A Peace, V Mcgilligan, I Bot
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Dutch Heart Foundation Background/Introduction Atherosclerosis is the underlying pathology of many cardiovascular diseases and is characterized by chronic inflammation in the larger arteries. Activation of the NLRP3 inflammasome is one of the drivers of inflammation during atherosclerosis. Purpose Therefore, we aimed to determine the ability of the novel bispecific anti-NLRP3 inflammasome antibody, called InflamAb, to inhibit atherosclerosis. Methods and Results In vitro treatment of bone marrow derived macrophages with 25 ng/mL InflamAb effectively inhibited the IL-1β release induced by NLRP3 inflammasome activation with LPS and Aluminium hydroxide (P&amp;lt;0.05). In addition, InflamAb administration in western-type diet fed Apoe-/- mice significantly reduced circulating IL-1β at 4 hours post inflammasome activation (P&amp;lt;0.05), thereby showing in vivo efficacy of InflamAb. Subsequently, we assessed the effect of InflamAb on both the development of atherosclerosis and on pre-existing plaques. Treatment of western-type diet fed female Apoe-/- mice with 100 µg InflamAb or isotype control antibody (3x per week i.p) significantly inhibited collar-induced atherosclerotic plaque development in the carotid artery from 59±8*10^3 µm^2 in control mice to 36±5*10^3 µm^2 (P&amp;lt;0.05), which coincided with a reduction in relative macrophage (control: 36±2% versus InflamAb: 28±3%, P&amp;lt;0.05) and necrotic core content (control: 16±2% versus InflamAb: 8±1%, P&amp;lt;0.05). InflamAb treatment of male Apoe-/- with pre-existing atherosclerosis did not affect lesion size, but improved plaque stability parameters as illustrated by a reduced relative macrophage (control: 48±2% versus InflamAb: 42±2%, P&amp;lt;0.05) and necrotic core (control: 21±1% versus InflamAb: 18±1%, P&amp;lt;0.05) content. In addition, we observed a trend towards increased collagen levels upon InflamAb treatment (control: 39±2% versus InflamAb: 44±2%, P=0.08). Conclusion To conclude, NLRP3 inflammasome inhibition by the bispecific antibody InflamAb shows promising efficacy in inhibiting atherosclerotic plaque development and destabilization.
Dissecting signal transduction pathways that regulate cardiomyocyte proliferation
Authors: R Artioli, R Cannilla, M Dal Ferro, L Zentilin, M Giacca, C Collesi
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements None. Introduction During cardiac development, the Notch, Hippo, and Wnt/β-catenin signalling pathways play a crucial role in the tuning of cardiomyocyte proliferation. The individual engagement of each of these pathways during the early postnatal life, however, is still unclear. Purpose We wanted to develop a method to visualize over time, both in vitro and in vivo, the topography of activation of the Notch, Wnt/β-catenin, and Hippo pathways and their crosstalk upon cardiomyocyte treatment with a series of pro-proliferative stimuli. Methods We obtained Adeno-Associated Virus (AAV) constructs expressing distinct fluorescent proteins (ECFP, GFP, mCherry, DsRed) under the control of promoters that are specifically activated downstream of the signalling cascades of interest (a synthetic promoter consisting of 12 CSL binding sites for Notch1, a synthetic promoter consisting of 8 TEAD binding sites for YAP, and a synthetic promoter containing 7 TCF/LEF1 binding sites for β-catenin). Results First, by high content microscopy we followed the time of the activation of the AAV vectors coding for fluorescent reporters, either alone or in combination. We observed that a crosstalk exists between Yap and Notch signalling in proliferating cardiomyocytes, as detected by the activation of their respective fluorescent reporters. We confirmed our data by gene expression analysis and co-immunoprecipitation using specific antibodies in fractionated cardiomyocyte extracts. Next, we observe that miR-1825, miR-199a-3p, and miR-590-3p – three pro-regenerative miRNAs in cardiomyocytes– specifically activate both Yap and Notch signalling. Finally we characterized the reporter systems in vivo, to dissect the intracellular signalling pathways that are active in the first week of postnatal life. Beta-catenin signalling was quite undetectable in any of the conditions tested. In contrast, both Notch and Yap showed significant activation, consistent with their role in regulating early postnatal cardiomyocyte proliferation. Conclusions The developed AAV-fluorescent reporters allow to efficiently and specifically visualize the pro-proliferative activation of Notch and Yap signalling both in vitro and in vivo. The results obtained are supportive of a crosstalk existing between Notch and Yap pathways in proliferating cardiomyocytes.
2-Aminoethoxydiphenyl borate prolongs atrial and shortens ventricular action potential duration through modulation of ion channels in the sarcolemma
Authors: M Rivaud, F M Bosada, S C M Van Amersfoort, C A Schumacher, R F Al Shama, J R De Groot, A O Verkerk, R F Coronel, M W Veldkamp
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): PPP allowance, Health holland ZonmW Background 2-Aminoethoxydiphenyl Borate (2-APB) has been shown to prevent atrial fibrillation in animal models, supposedly through its inhibitory action on the intracellular IP3 receptor and subsequent calcium regulation. However, 2-APB may modulate other ion channels or transporters and the cellular mechanisms associated with the prevention of atrial fibrillation are not well established. We aimed at unravelling the cellular effects of 2-APB on atrial and ventricular action potential and currents. Methods Rabbit (New Zealand White) left atrial and ventricular cardiomyocytes were isolated from Langendorff-perfused hearts by enzymatic dissociation. Adult human atrial cardiomyocytes were obtained by enzymatic dissociation from left atrial appendages (LAA) routinely removed from AF patients undergoing minimally invasive surgical pulmonary vein isolation. Action potentials (APs) and K+ currents were recorded at 36.5 °C with the amphotericin-B-perforated or whole cell patch clamp technique. RNA was isolated from left atrial and ventricular tissue and sequenced on an Illumina HiSeq4000 platform. Results In rabbit atrial cardiomyocytes, extracellular application of 5 uM 2-APB had no effect on action potential duration (APD), but extracellular application of 50 uM 2-APB prolonged the APD by 40% and inhibited an outward K+-current. Inversely, extracellular application of 5 uM 2-APB shortened the APD of rabbit left ventricular cardiomyocytes by 20% while extracellular application 50 uM 2-APB did not prolong the APD. Additionally, 2-APB did not modulate K+ currents in ventricular rabbit cardiomyocytes. Intracellular application (in the patch pipet) of 50 uM 2-APB did not have any effects on atrial APD, suggesting that IP3 receptor inhibition is not involved. However, subsequent application of 50 uM 2-APB in the bath prolonged the APD by 40% indicating that 2-APB prolongs atrial APD by inhibiting one or more K+ channel types in the sarcolemmal membrane from the extracellular site. RNA sequencing analysis revealed potential targets channel including TRP and Kv channels differentially expressed in atria versus ventricle. In human atrial cardiomyocytes from AF patients, application of 50 uM 2-APB prolonged APD by 25%. Conclusions 2-APB prolongs rabbit and human atrial APD and shortens rabbit ventricular APD in a concentration dependent manner, most likely due to modulation of sarcolemmal K+ channels, differentially expressed in the two cell types. Pharmacological studies with 2-APB may reveal novel cellular mechanisms identifying potential new targets for the treatment of atrial fibrillation.
Exploiting spatial transcriptomics to investigate the molecular mechanisms involved in cardiac metastasis development
Authors: D Lorizio, M Chiesa, M Pinamonti, R Bussani, G Ciucci, R Vuerich, S Zacchigna
Journal: Cardiovascular Research (2024)

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): IEO-Monzino Foundation Background Cardiac metastasis is considered a relatively rare malignancy, although an increasing prevalence is expected in the future due to the increasing life expectancy. Several cases of cardiac metastases remain untreatable, given the anatomical location that often hampers both surgical eradication and bioptic collection for biological characterization. Here, we use spatial transcriptomics to provide an unbiased molecular characterization of cardiac metastases, and identify novel targets for their treatment. Material and methods We performed spatial transcriptomics on samples of primary tumor, cardiac, and extra-cardiac metastasis from three different patients. First, we checked RNA integrity by means of DV200 analysis. Spatial transcriptomic analysis was performed by using the GeoMX technology (Nanostring), which provides whole-transcriptome data along with spatial distribution information of the preselected samples from the candidate regions of interest (ROIs). ROIs were selected on the formalin-fixed paraffin-embedded sections by means of geometric shapes in combination with cell markers specific for tumor cells, cardiomyocytes and endothelial cells, respectively. Differential gene expression analysis and Gene Set Enrichment Analysis (GSEA) were performed on the entire transcriptome. Cell-cell interactions between cardiac cells and cancer cells were then evaluated by means of InterCellaR. Results Differential expression analysis between cardiac metastases and extra-cardiac tumors (primary tumors + extra-cardiac metastases) identified 92 differentially expressed genes (DEGs). Of these, 53 genes were significantly overexpressed in cardiac metastases, while 39 genes were overexpressed in the extra-cardiac samples, regardless of the site of origin of the tumor. Unsupervised hierarchical clustering of protein-coding genes clearly separated the samples according to their anatomical location, in particular cardiac metastasis from extra-cardiac tumors. GSEA identified several pathways enriched in cardiac metastases compared to the extra-cardiac tumors, including mechanotransduction (e.g. keratinization and cell-matrix interaction) and triglyceride metabolism, suggesting a potential role of these signaling pathways in the development of cardiac metastasis. Interactome analysis revealed a rich plethora of receptor-ligand interactions in cardiac metastasis, especially between tumor cells and cardiomyocytes. Conclusions These results start shedding light on the mechanisms that allow cancer cell growth in the heart and pave the way to the development of novel therapies to treat this malignancy.