Citations

Guideline-recommended screening programs exist for only a few cancer types. Although all these programs are understood to lead to reductions in cancer-related mortality, standard-of-care screening tests vary in accuracy, adherence and effectiveness. Recent advances in high-throughput technologies and machine learning have facilitated the development of blood-based multi-cancer cancer early detection (MCED) tests. MCED tests are positioned to be complementary to standard-of-care screening and they may broaden screening availability, especially for individuals who are not adherent with current screening programs and for individuals who may harbor cancers with no available screening options. In this article, we outline some key features that should be considered for study design and MCED test development, provide an example of the developmental pathway undertaken for an emerging multi-biomarker class MCED test and propose a clinical algorithm for an imaging-based diagnostic resolution strategy following MCED testing.

Spinocerebellar ataxia type 3 (SCA3) is the most common type of disease related to poly-glutamine (polyQ) repeats. Its hallmark pathology is related to the abnormal accumulation of ataxin 3 with a longer polyQ tract (polyQ-ATXN3). However, there are other mechanisms related to SCA3 progression that require identifying trait and state biomarkers for a more accurate diagnosis and prognosis. Moreover, the identification of potential pharmacodynamic targets and assessment of therapeutic efficacy necessitates valid biomarker profiles. The aim of this review was to identify potential trait and state biomarkers and their potential value in clinical trials. Our results show that, in SCA3, there are different fluid biomarkers involved in neurodegeneration, oxidative stress, metabolism, miRNA and novel genes. However, neurofilament light chain NfL and polyQ-ATXN3 stand out as the most prevalent in body fluids and SCA3 stages. A heterogeneity analysis of NfL revealed that it may be a valuable state biomarker, particularly when measured in plasma. Nonetheless, since it could be a more beneficial approach to tracking SCA3 progression and clinical trial efficacy, it is more convenient to perform a biomarker profile evaluation than to rely on only one.

ImportanceCutaneous squamous cell carcinoma (CSCC) is the second most common malignant disease in the US. Although it typically carries a good prognosis, a subset of CSCCs are highly aggressive, carrying regional and distant metastatic potential. Due to its high incidence, this aggressive subset is responsible for considerable mortality, with an overall annual mortality estimated to equal or even surpass melanoma. Despite this morbidity, CSCC is excluded from national cancer registries, making it difficult to study its epidemiology and outcomes. Therefore, the bulk of the CSCC literature is composed of single-center and multi-institutional retrospective cohort analyses. Given variations in reporting measures and analyses in these studies, interpretability between studies and the ability to pool results are limited.ObjectiveTo define standardized reporting measures for retrospective CSCC studies.FindingsAn expert panel was convened to determine standardized guidelines for recording and analyzing retrospective CSCC data. A total of 13 dermatologists and dermatologic surgeons with more than 5 years of posttraining experience and considerable experience with performing CSCC outcomes research were recruited to the panel. Consensus recommendations were achieved for CSCC retrospective study reporting measures, definitions, and analyses.Conclusions and RelevanceThe recommendations in this report present the potential to standardize future CSCC retrospective studies. With such standardization, future work may have greater interstudy interpretability and allow for pooled analyses.

[ 18 F]-Flortaucipir positron emission tomography (PET) is considered a good biomarker of Alzheimer’s disease. However, it is unknown how flortaucipir is associated with the distribution of tau across brain regions and how these associations are influenced by amyloid-β. It is also unclear whether flortaucipir can detect tau in definite primary age–related tauopathy (PART). We identified 248 individuals at Mayo Clinic who had undergone [ 18 F]-flortaucipir PET during life, had died, and had undergone an autopsy, 239 cases of which also had amyloid-β PET. We assessed nonlinear relationships between flortaucipir uptake in nine medial temporal and cortical regions, Braak tau stage, and Thal amyloid-β phase using generalized additive models. We found that flortaucipir uptake was greater with increasing tau stage in all regions. Increased uptake at low tau stages in medial temporal regions was only observed in cases with a high amyloid-β phase. Flortaucipir uptake linearly increased with the amyloid-β phase in medial temporal and cortical regions. The highest flortaucipir uptake occurred with high Alzheimer’s disease neuropathologic change (ADNC) scores, followed by low-intermediate ADNC scores, then PART, with the entorhinal cortex providing the best differentiation between groups. Flortaucipir PET had limited ability to detect PART, and imaging-defined PART did not correspond with pathologically defined PART. In summary, spatial patterns of flortaucipir mirrored the histopathological tau distribution, were influenced by the amyloid-β phase, and were useful for distinguishing different ADNC scores and PART.

Background: Studies suggest circulating iron levels play a role in the pathophysiology of abdominal aortic aneurysms (AAAs). Low iron levels lead to medial degeneration and VSMC dysfunction while elevated iron levels promote oxidative stress. Hepcidin is a major iron regulatory hormone that binds ferroportin to reduce serum iron levels, and its removal in VSMCs has been shown to exacerbate AAAs; however, the role of sex in this mechanism remains understudied. The goal of the current study is to investigate the potential role of sex in iron metabolism in AAAs. Methods: Male and female C57BL/6 wild-type (WT/WT), heterozygous hepcidin knock-out (WT/KO) and hepcidin KO (KO/KO) underwent peri-adventitial application of porcine pancreatic elastase on the infrarenal aorta (n=10-19/group). At day 14, the aortas were measured using video micrometry and harvested for immunohistochemistry. In separate studies, male and female hepcidin WT/WT and KO/KO underwent elastase application and systemic b-aminopropionitrile (BAPN, 0.02%) administration and were harvested at 28 days. Finally, male and female LysM Cre Ferroportin flx/flx mice and their controls underwent elastase application and harvested at 14 days (n=5-10/group). Results: Hepcidin WT/KO and KO/KO male mice had dose dependent protection of aneurysm formation compared with controls (WT/WT: 145.5±33.8% vs WT/KO: 110.7±34.8% vs KO/KO: 53.7±13.7%, p=0.0001 by ANOVA). Female Hepcidin WT/KO and KO/KO mice exhibited increased AAA size compared with their controls (KO/KO: 119.0±23.5% vs WT/KO: 101.6 ±28.6% vs WT/WT: 71.2±22.0%, p =.0011 by ANOVA). In the elastase + BAPN model, Hepcidin KO/KO male mice had less aortic dilation compared to controls (KO/KO: 196.0 ±31.9% vs WT/WT: 476.8±82.2%, p =.0002 by Mann-Whitney) while female KO/KO mice exhibited greater aortic dilation (WT/WT: 186.7±26.0% vs. KO/KO: 406.9±62.2%, p=0.006). Male LysM Cre Ferroportin flx/flx mice had decreased aortic dilation (WT/WT: 132.5±19.3% vs flx/flx: 53.5±7.8%, p=0.001). Conclusion: Elimination of hepcidin in AAA formation exhibited protective effects in male and deleterious effects in female mice. This data suggests targeting of hepcidin using drug-based therapies could have gender-specific clinical implications.

The CD40-CD40L co-stimulatory dyad has been identified as critical player in atherosclerosis. Our previous work has shown that CD40 and CD40L have cell divergent effects on atherosclerosis. Deficiency of the CD40-CD40L dendritic cell-T cell axis decreased atherosclerosis via reducing T helper 1 responses, whereas deficiency of macrophages CD40 reduced atherosclerosis by enhancing efferocytosis and reducing necrotic core content. CD40 is highly expressed on B cells and interacts with CD40L on T cells to induce antibody production and T follicular helper cell responses. We therefore hypothesize that deletion of CD40 will reduce atherosclerosis. We aim to unravel the role of B cell CD40 in atherosclerosis. CD40 expression on B cell subsets was determined on peripheral blood mononuclear cells of patients and correlated with severity of coronary artery disease (CAD). Atherosclerosis was analyzed in CD19 Cre CD40 flfl ApoE -/- mice that were fed an atherogenic diet for 14 weeks. Patients with CAD showed a decrease in CD40 on their putative B1 cells, which was associated with increased plaque burden and decreased plaque fibrosis as detected by coronary intravascular ultrasound. Depletion of CD40 on B cells in ApoE -/- mice resulted in a decreased germinal center formation and inadequate IgM production as expected. However, in line with our patient data, absence of B cell-CD40 increased atherosclerosis. This was caused by a reduction in B1 cells and consequently a reduction in atheroprotective anti-oxidation specific epitope (OSE) IgMs. Transfer of CD40-competent B1b cells in B cell-CD40-deficient mice restored anti-OSE IgM levels and prevented the increase in atherosclerosis. CD40-deficient B1b cells have an altered mTOR signaling pathway, impaired mitochondrial function, excessive uptake of lipids, increased cellular stress and accelerated cell death. Data indicate that CD40-deficiency promotes phosphorylation of RAPTOR and protein kinase B (AKT). We have identified a novel function of CD40 on B1b cells. B1b cell-CD40 exerts an anti-atherogenic role by regulating B1b cell metabolic homeostasis via mTOR pathway. Further mechanistic insights into how CD40 mounts adequate atheroprotective anti-OSE IgM production are currently being investigated.

Objective: We recently discovered that activation of oxidized soluble guanylate cyclase with ataciguat (ATA) slows valve calcification and dysfunction in mice and humans with moderate aortic valve stenosis (AVS). We examined if ATA would attenuate multiple mechanisms contributing to AVS and end organ dysfunction associated with AVS, and hypothesized: 1) ATA attenuates osteogenic signaling in valve tissue from mice with AVS, thereby attenuating calcification and valve dysfunction, 2) AVS increases rates of sudden cardiac death in our model, and 3) treatment with ATA prevents QT prolongation and deleterious electrical remodeling commonly observed in AVS. Methods: Functional, molecular, and histopathological changes in valve tissue along with QT interval duration were measured in 3 independent groups of mice: non-AVS, AVS, and AVS+ATA (WD for 6 months then WD+ATA for 3 months (AVS+ATA). We gave 5 mg/kg isoproterenol (ISO, i.p.) to ldlr -/- /ApoB100-only mice fed normal chow (non-AVS) or western diet (AVS, fed WD for 9 months) to assess risk for malignant arrhythmias. Retrospective analysis of QT interval was also done on data from our recent Phase 1 clinical trial. Results: AVS+ATA mice had less valve calcification and slower progression of valve dysfunction through attenuation of canonical BMP signaling and reductions in Runx2 and SPP1 compared to AVS mice. QT prolongation was evident in AVS mice, and was significantly attenuated by treatment with ATA (p= 0.001) and coarsely associated with severity of valvular or ventricular function. While all non-AVS mice survived for 24 hours after ISO, 40% of mice with AVS reached a moribund equivalent within 4 hours. Retrospective analyses also showed reductions in QT interval duration in patients treated with 200 mg/day ATA for 14 days (p < 0.05). Conclusion: Collectively, this is the first study reporting increased propensity for malignant arrhythmias in our mouse model of AVS, QT prolongation as a likely underlying mechanism, and restoration of cardiac conduction dynamics with targeted restoration of nitric oxide signaling. Future studies focused on the role of sGC in the evolution and prevention of AVS and consequent deleterious electrical remodeling/malignant arrhythmia risk is warranted.

The accumulation of senescent cells, a hallmark phenotype of aging tissues, elevates the risk of cardiovascular disease with age. Our prior studies have illustrated a crucial role played by senescent cells in promoting coagulation during murine atherosclerosis through the senescence-associated secretory phenotype (SASP). To explore the heterogeneity of senescent vascular cells in thrombotic vascular diseases and identify potential druggable pathways to counter senescence, we utilized the senescence reporter mouse p16TdTomato+/-, which was injected with an adeno-associated virus expressing Proprotein Convertase Subtilisin/Kexin type 9 to induce atherosclerosis when subjected to a high-fat diet (HFD). In a subset of HFD mice, some were left untreated, while others were treated with senolytic drug ABT-737 to selectively eliminate senescent cells. Aortas from ABT-737-treated mice exhibited increased collagen deposition and reduced arterial stiffness. Paired with pseudo-bulk RNA-sequencing analysis, we observed that HFD-induced extracellular matrix (ECM) remodeling genes were diminished by ABT-737, suggesting that ECM-platelet interactions may be influenced by senescence. Single-cell RNA-sequencing analysis from aortas identified 28 distinct cell clusters. Utilizing Gene Set Enrichment Analysis with the SenMayo and CellAge panels, we identified four cell clusters exhibiting increased senescent features that were mitigated by ABT-737. These included two VSMC clusters, fibroblasts, and neutrophil-like cells. Unbiased subclustering of potentially senescent clusters revealed a small population of cells enriched with HFD and reduced by ABT-737, confirming their senescent nature. In the most abundant cluster, Spp1 and Serpine2 strongly represented the senescent VSMC subpopulation, enriched for TGFβ signaling, ECM remodeling, and coagulation cascade genes. Further, we validated these findings through spatial transcriptomics and single-molecule FISH of brachiocephalic arteries from atherosclerotic mice and atherosclerotic mice depleted of senescent cells. Our results reveal a novel transcriptomic signature for senescent vascular cells that holds potential for therapeutic targeting in age-related vascular diseases.