Preclinical Phase Research Articles

Heparin interferes with the uptake of liposomes in glioma

In glioblastoma, a malignant primary brain tumor, liposomes have shown promise in pre-clinical and early phase clinical trials as delivery vehicles for therapeutics. However, external factors influencing cellular uptake of liposomes in glioma cells are poorly understood. Heparin and heparin analogues are commonly used in glioma patients to decrease the risk of thrombo-embolic events. Our results show that heparin inhibits pegylated liposome uptake by U87 glioma and GL261 cells in a dose dependent manner in vitro, and that heparin-mediated inhibition of uptake required presence of fetal bovine serum in the media. In a subcutaneous model of glioma, Cy5.5 labeled liposomes could be detected with in vivo imaging after direct intra-tumoral injection. Ex-vivo analysis with flow cytometry showed a decreased uptake of liposomes into tumor cells in mice treated systemically with heparin compared to those treated with vehicle only.

Developing Vaccines to Improve Preparedness for Filovirus Outbreaks: The Perspective of the USA Biomedical Advanced Research and Development Authority (BARDA).

Outbreaks of viral hemorrhagic fever caused by filoviruses have become more prevalent in recent years, with outbreaks of Ebola virus (EBOV), Sudan virus (SUDV), and Marburg virus (MARV) all occurring in 2022 and 2023. While licensed vaccines are now available for EBOV, vaccine candidates for SUDV and MARV are all in preclinical or early clinical development phases. During the recent outbreak of SUDV virus disease, the Biomedical Advanced Research and Development Authority (BARDA), as part of the Administration for Strategic Preparedness and Response within the U.S. Department of Health and Human Services, implemented key actions with our existing partners to advance preparedness and enable rapid response to the outbreak, while also aligning with global partners involved in the implementation of clinical trials in an outbreak setting. Beyond pre-existing plans prior to the outbreak, BARDA worked with product sponsors to expedite manufacturing of vaccine doses that could be utilized in clinical trials. While the SUDV outbreak has since ended, a new outbreak of MARV disease has emerged. It remains critical that we continue to advance a portfolio of vaccines against SUDV and MARV while also expediting manufacturing activities ahead of, or in parallel if needed, outbreaks.

Application and validation of an algorithmic classification of early impairment in cognitive performance

Objective Due to the long prodromal period for dementia pathology, approaches are needed to detect cases before clinically recognizable symptoms are apparent, by which time it is likely too late to intervene. This study contrasted two theoretically-based algorithms for classifying early cognitive impairment (ECI) in adults aged ≥50 enrolled in the Baltimore Longitudinal Study of Aging. Method Two ECI algorithms were defined as poor performance (1 standard deviation [SD] below age-, sex-, race-, and education-specific means) in: (1) Card Rotations or California Verbal Learning Test (CVLT) immediate recall and (2) ≥1 (out of 2) memory or ≥3 (out of 6) non-memory tests. We evaluated concurrent criterion validity against consensus diagnoses of mild cognitive impairment (MCI) or dementia and global cognitive scores using receiver operating characteristic (ROC) curve analysis. Predictive criterion validity was evaluated using Cox proportional hazards models to examine the associations between algorithmic status and future adjudicated MCI/dementia. Results Among 1,851 participants (mean age = 65.2 ± 11.8 years, 50% women, 74% white), the two ECI algorithms yielded comparably moderate concurrent criterion validity with adjudicated MCI/dementia. For predictive criterion validity, the algorithm based on impairment in Card Rotations or CVLT immediate recall was the better predictor of MCI/dementia (HR = 3.53, 95%CI: 1.59–7.84) over 12.3 follow-up years. Conclusions Impairment in visuospatial ability or memory may be capable of detecting early cognitive changes in the preclinical phase among cognitively normal individuals.

An Introductory Qualitative Exploration of Medical Student Perceptions of Professionalism at One Medical University.

Professionalism as a competency in medical education has been defined in multiple ways. Irby and Hamstra offered three frameworks of professionalism in medical education. This study examines medical students' definitions of professionalism to assess whether they align with these frameworks. We administered an open-ended questionnaire to 92 medical students at a single university in the United States. We conducted thematic coding of responses and calculated code frequencies. The response rate was 54%. There were no observable differences between the responses of students in clinical versus preclinical training phases. The majority of comments (84%) reflected aspects of multiple frameworks from Irby and Hamstra and three emergent themes were identified. Most respondents (96%) cited aspects of the behavior-based framework. Most students' (66%) responses also aligned with the virtue-based framework. Emergent themes were "hierarchical nature of medicine," "academic environment/hidden curriculum," and "service and advocacy." "Service and advocacy" can be viewed as contexts for Irby and Hamstra's identity formation framework, but references did not align with the full definition. Our findings suggest that students view professionalism through multiple frameworks and indicate a predominance of the behavior-based framework. Experiences with organizational culture and values may be important in students' definitions of professionalism.

#4321 ACTION3 PHASE 3 STUDY: EVALUATING A NOVEL THERAPEUTIC STRATEGY OF A CCR2 INHIBITOR (DMX-200) WITH ANGIOTENSIN RECEPTOR BLOCKADE IN FSGS

Abstract Background and Aims Focal segmental glomerulosclerosis (FSGS) is a disease of podocytes. Complications include nephrotic syndrome and progressive kidney failure. FSGS is a condition with high unmet need and there is no approved treatment. The angiotensin II receptor type 1 (AT1R) and chemokine receptor 2 (CCR2) are G protein coupled receptors that form functional heteromers. Simultaneous antagonism of these receptors suggested synergistic renoprotective effects in preclinical and early phase clinical studies of proteinuric kidney disease. DMX-200 (repagermanium) is a C-C chemokine receptor type 2 (CCR2) inhibitor that, when administered concurrently with an angiotensin II receptor blocker (ARB), is designed to inhibit recruitment of monocytes implicated in the inflammatory chemokine environment of chronic disease. In a Phase 2a open-label study of 27 patients with proteinuric chronic kidney disease, 25% of patients achieved > 50% reduction in uPCR with combined use of DMX-200 and irbesartan. A Phase 2a placebo-controlled cross-over study in 8 patients with primary FSGS receiving stable dose of irbesartan demonstrated evidence of promising efficacy with clinically relevant reduction in uPCR of 17% with DMX-200 compared with placebo. These encouraging data suggest that treatment with DMX-200 may result in clinically meaningful reduction in proteinuria when added to ARB in patients with glomerular diseases such as FSGS. Proteinuria reduction is a positive prognostic sign for preserving kidney function. The observations from earlier trials have led to the initiation of an international randomised double-blind, placebo-controlled Phase 3 clinical trial (ACTION3) to further evaluate the efficacy of DMX-200 in patients with FSGS receiving an ARB. Method Eligible patients are adults (18-80 years) with biopsy-proven primary FSGS, genetic FSGS or FSGS of undetermined cause (FSGS-UC), and uPCR of >1.5g/g based on 24hr urine collection and eGFR of ≥25 ml/min/1.73 m2. Blood pressure ≤160/100 mmHg and BMI ≤40kg/m2. Patients with secondary FSGS or those treated with strong immunomodulatory agents (other than corticosteroids) are excluded. All patients are required to receive concomitant ARB treatment at a dose of at least 50% of the maximum approved dose according to the relevant product label. Patients receiving ACE-inhibitors are required to switch to an ARB. Other background therapies such as SGLT-2 inhibitors and mineralocorticoid receptor antagonists are permitted if receiving a stable dose in the 3 months prior to screening. Patients are randomized in a 1:1 ratio to DMX-200 (120 mg twice daily) or placebo. The primary objective is to evaluate the efficacy of DMX-200 on reduction in PCR after 35 weeks of treatment and eGFR slope after 104 weeks of treatment. Secondary objectives are safety and tolerability of DMX-200 and to evaluate the effect of DMX-200 on response criteria and composite endpoint of worsening kidney function. Exploratory objectives include assessment of changes in inflammatory biomarkers such as MCP-1 and the pharmacokinetic profile of DMX-200. Two interim analyses (IA) are planned, first planned IA for futility will be performed by an Independent Data Monitoring Committee after the first ​72 randomized patients (36 patients per group) complete 35 weeks ​of treatment. A second IA will be performed once approximately​ 144 enrolled patients complete 35 weeks ​of treatment.​ The final analysis will be performed once approximately 286 patients complete 104 weeks of treatment. The study is currently open in 11 countries at approximately 75 investigational sites and will be expanded to include more countries and sites subject to a successful first interim analysis. Results ACTION3 is a trial in progress and is expected to be completed in 2026. Conclusion ACTION3 is a Phase 3 randomized, placebo-controlled efficacy and safety trial which evaluates the novel approach of combining an investigational CCR2 inhibitor, DMX-200, with angiotensin receptor blockade in patients with FSGS.

Oral Maxillofacial Surgeon – A Hidden Weapon in Head and Neck Oncology

Oral squamous cell carcinoma dominates all the oral cancer cases with potentially malignant disorders which are also recognised as detectable pre-clinical phase of oral cancer. Patients with head and neck cancer should get a thorough dental oncologic examination as part of their pre-treatment evaluation. Dental professionals that specialise in oral and maxillofacial (OMF) surgery are the best people to manage patients with tumours in this area. This study discusses the significance of having these oncology-trained surgeons on the head and neck surgical teams, their underappreciated contributions to raising awareness of oral cancer and its negative effects among the general public as well as teaching oral cancer patients proper oral hygiene techniques. It also discusses the difficulties they face today to practise oncosurgery and be respected by their medical colleagues and provides an answer to the question of whether OMF surgeons have any role in such surgical setups.

Development and qualification of cell-based relative potency assay for a human respiratory syncytial virus (RSV) mRNA vaccine

Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract infections worldwide. A safe and effective RSV vaccine has been an elusive goal but recent advances in vaccine technology have improved the likelihood that a vaccine for the prevention of RSV could be licensed in near future. We have developed an RSV vaccine V171 consisting of four lipids and messenger ribonucleic acid (mRNA) encoding an engineered form of the RSV F protein stabilized in its prefusion conformation. The lipids form lipid nanoparticles (LNP) with mRNA encapsulated during process, which protects the mRNA from degradation and enables the mRNA to be delivered into mammalian cells. Once inside the cells, the mRNA then can be translated into RSV F protein and elicit both humoral and cellular immune responses. Preclinical results and Phase I clinical trial results indicate that this mRNA vaccine targeting RSV F protein is a promising RSV vaccine approach and should be further evaluated in clinical trials. We have developed a cell-based relative potency assay to support the Phase II development of this vaccine. Test articles and a reference standard are tested with serial dilutions in a 96-well plate pre-seeded with Hep G2 cells. Cells were incubated for 16–18 h after transfection and then permeabilized and stained with a human monoclonal antibody specific to RSV F protein, followed by a fluorophore-conjugated secondary antibody. The plate is then analyzed for percentage of transfected cells and relative potency of the test article is calculated by comparing its EC50 to that of a reference standard. This assay takes advantage of the fact that due to the inherent variability in biological test systems an absolute measure of potency is more variable than a measure of activity relative to a standard. Targeting testing relative potency range 25–250 %, our assay showed an R2 close to 1 for linearity, relative bias of 1.05–5.41 %, and intermediate precision of 11.0 %. The assay has been used for testing of process development samples, formulation development samples, as well as drug product intermediate (DPI) and drug product (DP) in support of Phase II development of our RSV mRNA vaccine.

Longitudinal blood biomarker trajectories in preclinical Alzheimer's disease.

Plasma biomarkers are altered years prior to Alzheimer's disease (AD) clinical onset. We measured longitudinal changes in plasma amyloid-beta (Aβ)42/40 ratio, pTau181, pTau231, neurofilament light chain (NfL), and glial fibrillary acidic protein (GFAP) in a cohort of older adults at risk of AD (n=373 total, n=229 with Aβ and tau positron emission tomography [PET] scans) considering genetic and demographic factors as possible modifiers of these markers' progression. Aβ42/40 ratio concentrations decreased, while NfL and GFAP values increased over the 4-year follow-up. Apolipoprotein E (APOE) ε4 carriers showed faster increase in plasma pTau181 than non-carriers. Older individuals showed a faster increase in plasma NfL, and females showed a faster increase in plasma GFAP values. In the PET subsample, individuals both Aβ-PET and tau-PET positive showed faster plasma pTau181 and GFAP increase compared to PET-negative individuals. Plasma markers can track biological change over time, with plasma pTau181 and GFAP markers showing longitudinal change in individuals with preclinical AD. Longitudinal increase of plasma pTau181 and glial fibrillary acidic protein (GFAP) can be measured in the preclinical phase of AD. Apolipoprotein Eε4 carriers experience faster increase in plasma pTau181 over time than non-carriers. Female sex showed accelerated increase in plasma GFAP over time compared to males. Aβ42/40 and pTau231 values are already abnormal at baseline in individuals with both amyloid and tau PET burden.

Normative data for COGITAB: An Italian tablet-based test battery conceived for the preclinical phase of Alzheimer’s disease

The number of people with dementia is increasing worldwide. Two main approaches have been adopted to identify subjects with Alzheimer’s disease (AD): the neuropsychological evaluation and the identification of biomarkers of AD. The first method is less invasive and easier to perform. This study assesses the psychometric properties of COGITAB, a novel web application d esigned to be sensitive to the subtle cognitive changes distinctive of the early Mild Cognitive Impairment (MCI) and the preclinical phase of AD. We enrolled 518 healthy controls, classified according to several risk factors and the presence of a family history of dementia. The participants were given COGITAB after a neuropsychological screening. The COGITAB Total Score (TS) was significantly affected by age and years of education. Acquired risk factors and family history of dementia significantly impacted only the COGITAB total execution time (TET), not the TS. This study provides normative data for a newly developed web application. Control subjects with acquired risk factors performed slower, giving an important role to the TET recording. Further studies should examine the ability of this new technology to discriminate between healthy subjects and subjects with initial cognitive decline, even when not detected by standard neuropsychological assessments.

Increased gut permeability and intestinal inflammation precede arthritis onset in the adjuvant-induced model of arthritis

BackgroundIntestinal inflammation, dysbiosis, intestinal permeability (IP), and bacterial translocation (BT) have been identified in patients with spondyloarthritis but the time at which they appear and their contribution to the pathogenesis of the disease is still a matter of debate.ObjectivesTo study the time-course of intestinal inflammation (I-Inf), IP, microbiota modification BT in a rat model of reactive arthritis, the adjuvant-induced arthritis model (AIA).MethodsAnalysis was performed at 3 phases of arthritis in control and AIA rats: preclinical phase (day 4), onset phase (day 11), and acute phase (day 28). IP was assessed by measuring levels of zonulin and ileal mRNA expression of zonulin. I-inf was assessed by lymphocyte count from rat ileum and by measuring ileal mRNA expression of proinflammatory cytokines. The integrity of the intestinal barrier was evaluated by levels of iFABP. BT and gut microbiota were assessed by LPS, soluble CD14 levels, and 16S RNA sequencing in mesenteric lymph node and by 16S rRNA sequencing in stool, respectively.ResultsPlasma zonulin levels increased at the preclinical and onset phase in the AIA group. Plasma levels of iFABP were increased in AIA rats at all stages of the arthritis course. The preclinical phase was characterized by a transient dysbiosis and increased mRNA ileal expression of IL-8, IL-33, and IL-17. At the onset phase, TNF-α, IL-23p19, and IL-8 mRNA expression were increased. No changes in cytokines mRNA expression were observed at the acute phase. Increased CD4+ and CD8+ T cell number was measured in the AIA ileum at day 4 and day 11. No increase in BT was observed.ConclusionThese data show that intestinal changes precede the development of arthritis but argue against a strict “correlative” model in which arthritis and gut changes are inseparable.

Preclinical Characterization and Phase 1 Trial Results of INBRX-109, a Third-Generation, Recombinant, Humanized, Death Receptor 5 Agonist Antibody, in Chondrosarcoma.

Patients with unresectable/metastatic chondrosarcoma have poor prognoses; conventional chondrosarcoma is associated with a median progression-free survival (PFS) of <4 months after first-line chemotherapy. No standard targeted therapies are available. We present the preclinical characterization of INBRX-109, a third-generation death receptor 5 (DR5) agonist, and clinical findings from a phase 1 trial of INBRX-109 in unresectable/metastatic chondrosarcoma (NCT03715933). INBRX-109 was first characterized preclinically as a DR5 agonist, with binding specificity and hepatotoxicity evaluated in vitro and antitumor activity evaluated both in vitro and in vivo. INBRX-109 (3 mg/kg every 3 weeks) was then evaluated in a phase 1 study of solid tumors that included a cohort with any subtype of chondrosarcoma and a cohort with IDH1/IDH2-mutant conventional chondrosarcoma. The primary endpoint was safety. Efficacy was an exploratory endpoint, with measures including objective response, disease control rate, and PFS. In preclinical studies, INBRX-109 led to antitumor activity in vitro and in patient-derived xenograft models, with minimal hepatotoxicity. In the phase 1 study, INBRX-109 was well tolerated and demonstrated antitumor activity in unresectable/metastatic chondrosarcoma. INBRX-109 led to a disease control rate of 87.1% (27/31; durable clinical benefit, 40.7% [11/27]), including 2 partial responses, and median PFS of 7.6 months. Most treatment-related adverse events, including liver-related events, were low grade (grade ≥3 events in chondrosarcoma cohorts, 5.7%). INBRX-109 demonstrated encouraging antitumor activity with a favorable safety profile in patients with unresectable/metastatic chondrosarcoma. A randomized, placebo-controlled, phase 2 trial (ChonDRAgon, NCT04950075) will further evaluate INBRX-109 in conventional chondrosarcoma.

Impact of APOE ε4 allele on associations between sub‐threshold, positive Amyloid‐β retention, brain function, and cognitive performances in cognitively normal older adults

AbstractBackgroundA growing body of evidence suggests a deteriorating effect of sub‐threshold amyloid‐beta (Aβ) accumulation on cognition before the onset of clinical symptoms of Alzheimer’s disease. Despite the association between the Aβ‐dependent pathway and the APOE ε4 allele, the impact of this allele on the progression from the sub‐threshold Aβ deposits to cognitive function impairment is unclear. Furthermore, the comparative analysis of positive Aβ accumulation in the preclinical phase is lacking. This study aimed to explore the differential effect of the APOE ε4 allele on the association between Aβ retention, brain function, and cognitive performance in cognitively healthy older adults, depending on the Aβ burden status.MethodOne hundred and eighty‐two older adults with normal cognition who underwent resting‐state functional magnetic resonance imaging were enrolled. They were dichotomized using [18F]‐labeled flutemetamol positron emission tomography into subjects with sub‐threshold (cognitively normal [CN] sub‐Aβ, n = 110; [APOE ε4 carrier, n = 30; non‐carrier, n = 80] and positive Aβ deposits [CN Aβ+ group, n = 72; APOE ε4 carrier, n = 34; non‐carrier, n = 38]).ResultWe found (i) distinct effects of the APOE ε4 allele and relationships with cognitive function in the preclinical phase; (ii) differences in local functional connectivity (FC) of the brain regions in networks vulnerable to Alzheimer’s disease according to the APOE ε4 allele, which was associated with higher executive and memory function; (iii) a differential association between regional Aβ retention and local FC according to the APOE ε4 allele; and (iv) a significant interaction between APOE ε4 allele and local FC for the remote inter‐network FC between the default‐mode and central‐executive network in the CN sub‐Aβ group, contributing a compensatory role for cognitive performance. Finally, these results were modulated according to Aβ positivity.ConclusionThis study is the first attempt to thoroughly examine the mechanism at play in the earliest phase of Alzheimer’s disease, focusing on the influence of the APOE ε4 allele from the sub‐threshold to positive Aβ retention during the preclinical phase.

How to Prevent and/or Revert Alzheimer's Disease Continuum During Preclinical Phases.

The development of Alzheimer's disease (AD) follows three consecutive phases: namely preclinical, prodromal or mild cognitive impairment (MCI), and dementia. In addition, the preclinical phase can be divided into subphases related to the presence of biomarkers that appear at different points before the onset of MCI. Indeed, an early risk factor could promote the appearance of additional ones through a continuum. The presence of various risk factors may trigger specific biomarkers. In this review, we comment on how modifiable risk factors for AD may be reverted, thus correlating with a possible decrease in the specific biomarkers for the disease. Finally, we discuss the development of a suitable AD prevention strategy by targeting modifiable risk factors, thereby increasing the level of "precision medicine" in healthcare systems worldwide.

The safety and efficacy of SNK01 (autologous natural killer cells) in combination with cytotoxic chemotherapy after failure of prior tyrosine kinase inhibitor in non-small cell lung cancer: Preclinical mouse model and phase I/IIa clinical study.

9057 Background: It is challenging to choose subsequent treatment option in patients with Osimertinib resistance in epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC). Meanwhile, antitumor effect of the antibody-dependent cellular cytotoxicity of natural killer (NK) cells mediated by cetuximab –anti-EGFR monoclonal antibody– has been reported. Therefore, we aimed to evaluate the safety and efficacy of SNK01 (non-genetically modified autologous NK cell) in combination with cytotoxic chemotherapy including cetuximab in NSCLC after failure to prior tyrosine kinase inhibitor (TKI) in a preclinical humanized cell line-derived xenograft (CDX) mouse model and phase I/IIa clinical study. Methods: We established a humanized CDX mouse model using Osimertinib-resistant lung cancer cell line. Reconstruction of the humanized mouse, CDX-related skills, and analytic data were made according to C&amp;SR Inc. manufacturing technique &amp; SOP protocol. The mice were divided into 4 groups based on treatment (no treatment [n = 2]; Cetuximab only [n = 3]; SNK01 only [n = 4]; SNK01 plus Cetuximab [n = 4]) and treated weekly for 5 weeks (SNK01, 1x107 cells/dose; Cetuximab, 20 ug/dose). In the clinical study, 12 patients with EGFR-mutated NSCLC who failed prior TKI treatment were finally enrolled. They received weekly SNK01 in combination with Gemcitabine/Carboplatin (n = 6) or Gemcitabine/Carboplatin/Cetuximab (n = 6), and dose escalation of SNK01 following “3+3” design (4×109 cells/dose or 6×109 cells/dose). The primary endpoint was safety, and secondary endpoint was efficacy. Results: In the preclinical study, flow cytometry analysis showed that NK cells (CD45+/CD56+/CD3-) were significantly increased in the groups administrated SNK01. The volume of tumor extracted after completion of treatment was the smallest in SNK01 plus cetuximab group. In the clinical study, the median age of patients was 61 years, 33.3% were male, and all patients had adenocarcinoma. The enrolled patients received weekly infusions of SNK01 for 7 to 8 weeks (4×109 cells/dose [n = 6]; 6×109 cells/dose [n = 6]). Since dose limiting toxicity was not observed, maximum tolerated dose of SNK01 was determined to be 6×109 cells/dose. SNK01-related adverse event ≥grade 3 was also not observed. In the efficacy analysis, objective response rate was 25%, disease control rate was 100% (partial response, n = 3/12; stable disease, n = 9/12), and median progression free survival (PFS) was 143 days. PFS will be updated as some patients are still being followed. Conclusions: SNK01 in combination with cytotoxic chemotherapy, including cetuximab in EGFR-mutated NSCLC with resistance to TKI was safe and showed a potential antitumor effect in this preclinical study and early phase I/IIa clinical trial. Clinical trial information: NCT04872634 .

Preclinical circulating tumor DNA (ctDNA) shedding duration and prognostic implications of modeling 3669 participants from American Cancer Society Cancer Prevention Study-3 (CPS-3) and Circulating Cell-free Genome Atlas substudy 3 (CCGA3).

3060 Background: Early detection of cancer occurs in the preclinical phase, when development of cancers is unobservable. Shedding of ctDNA has previously been associated with aggressive cancers, suggesting these tumors may develop rapidly before diagnosis. Assays of ctDNA in blood samples from prospective cohort studies at various lead times (interval between blood draw and diagnosis) may provide insight into the timescales of preclinical cancer development, as well as prognosis. Methods: We used a targeted methylation assay and the associated locked classifier for cancer signal detection (CSD) on samples from two large prospective biobank studies: 1) CPS-3, where blood draw at enrollment (2006-13) occurred ≤3 years (Ys) before clinical diagnosis (n=1065); 2) CCGA3 (NCT02889978), where blood draw (2016-19) was concurrent with clinical diagnosis (n=2604). We constructed a model for detectability including study, inherent shedding of ctDNA at clinical diagnosis, and effect of lead time by cancer type and stage. We additionally evaluated prognosis stratified by ctDNA status (CSD positivity), compared to expected cancer-specific survival (CSS) matched by age, cancer type, and stage for ≤9 Ys of follow-up, noting that ctDNA status may change during lead time. Results: Estimated inherent ctDNA detectability did not differ between CCGA3 and CPS-3 studies (95% confidence interval [CI] for difference is [-8.2%, 4.6%] and spans zero). Median [CI] detection timescales for exponential decay of detection by time were 0.75 [0.47, 1.3] Ys for localized stage, 0.89 [0.61, 1.33] Ys for regional, and 1.2 [0.84, 1.67] Ys for distant, consistent with a published model, and ranged by type from 0.49 [0.26, 0.88] Ys for pancreas to 2.45 [1.14, 4.87] for lymphoma. For CPS-3, estimated CSS at 9 Ys from blood draw was 54% for ctDNA+ and 79.5% for ctDNA- participants. Hazard ratios (HR) [CI] for ctDNA+ vs ctDNA- alone predicting CSS were 7 [4.2, 12.2], 4.2 [2.9, 6], and 3.5 [2.5, 4.7] for those diagnosed up to 1, 2, and 3 Ys after blood draw, respectively. Compared to synthetic controls for age, cancer type, stage, and lead time in Surveillance, Epidemiology, and End Results (SEER) data, CSS was better in each group: ctDNA+ HR 0.63 (CI: 0.54, 0.74); ctDNA- HR 0.45 (CI: 0.22, 0.74). Conclusions: These results provide estimates for the average detection window in tumors shedding ctDNA across multiple cancer sites and stages, including cancer types with no current screening paradigm. Taking lead time into account, preclinical ctDNA status selected clinically significant cancers, limiting potential for overdiagnosis. Additionally, long-term survival remained better than in SEER data, suggesting that ctDNA+ cancers can be survivable. Results require confirmation in future prospective analyses.

Quadratic associations between subthreshold amyloid‐beta retention and cortical volume modulated by APOE genotype in cognitively normal older adults

AbstractBackgroundA growing body of evidence suggests a deteriorating effect of subthreshold amyloid‐beta (Aβ) accumulation before the onset of clinical symptoms of Alzheimer’s disease (AD). Despite the association between the Aβ‐dependent pathway and the APOE ε4 allele, the modulating impact of this allele on the associations between subthreshold Aβ retention, cortical atrophy, and neuropsychological performance is unclear. This study aimed to explore the differential effect of the APOE ε4 allele on the association between subthreshold Aβ retention, cortical volume, and cognitive performance in cognitively healthy older adults (CN).MethodA total of 112 CN with subthreshold Aβ retention, consisting of 80 APOE ε4 non‐carrier and 32 APOE ε4 carrier, were included in the study. Participants underwent structural magnetic resonance imaging, [18F] flutemetamol (FMM) PET‐CT, and neuropsychological battery. Potential interactions between APOE status and Aβ accumulation were assessed with F‐tests implemented on both linear and quadratic interaction terms, adjusting for age, sex, and education years.ResultWe found a significant interaction between APOE ε4 allele, subthreshold Aβ deposits, cognitive function for cortical volume of predefined brain regions vulnerable to early Aβ retention. In CN with APOE ε4 carrier, regional Aβ retention in frontal, parietal, temporal lobe, and posterior cingulate cortex/precuneus showed a U‐shaped relationship with cortical volumes of superior parietal, fusiform gyrus, and amygdala. Furthermore, cortical volumes of superior, inferior temporal lobe, and parahippocampal gyrus in APOE ε4 carrier displayed a U‐shaped relationship with attention and executive function scores.ConclusionThis study is the first attempt to thoroughly examine the mechanism at play in the earliest phase of Alzheimer’s disease, focusing on the influence of the APOE ε4 allele on the association between subthreshold Aβ retention and cortical volume during the preclinical phase.

FRI-521 - SNP-630, a novel compound with multiple mechanisms, reverses liver inflammation and fibrosis in preclinical model and NASH phase 2 clinical trial

FRI-521 - SNP-630, a novel compound with multiple mechanisms, reverses liver inflammation and fibrosis in preclinical model and NASH phase 2 clinical trial

CARD24: Protective Effect of HemoTech - Blood Substitute Against Contrast Media-Induced Endothelial Toxicity: An In Vitro Study

Background: Although the most common complication after intravascular administration of iodinated contrast media (CM) is contrast-induced nephropathy (CIN) there is some evidence suggesting CM toxicity to endothelial cells (EC). CIN mechanism is well defined and includes medullary ischemia and direct toxicity to renal tubular cells. The mechanism of CM-associated EC damage is still unclear. The purpose of this study was twofold: (1) to determine the response of normal and ischemic coronary artery EC to low-osmolar nonionic CM commonly used in percutaneous coronary interventions (PCI) and (2) to evaluate the protective effect of HemoTech used concomitantly with CM. HemoTech is Hb-based blood substitute pharmacologically modified with vasodilators, anti-inflammatory and antioxidant agents: ATP, adenosine and reduced glutathione (GSH), and is in the preclinical development phase of its intended use as perfusion fluid for PCI. Methods: In this study human coronary artery EC (Clonetics, Bio-Whittaker, San Diego, CA) were subcultured in EGM-MV medium to confluence on cover slips and 6-well cell cultured plates in a humidified atmosphere of 5% CO2 and 37 deg C. To mimic ischemia, EC were depleted from GSH by 24 hr treatment with 200 uM of BSO (L-buthionine-[S,R]-sulphoximine).Then, normal and ischemic EC were exposed for 1, 2, 4 and 18 hrs to 50 uL/mL of low-osmolar nonionic iodinated CM Optiray 350 (Ioversol Injection 74%, Mallinckrodt Pharmaceuticals) alone or with 0.4 mM of HemoTech and assessed for cellular redox state and oxidative stress (GSH, TBARS), inflammatory response (VCAM-1) and early and late apoptosis. Results: The results demonstrate a different response of normal and ischemic EC to CM. While normal EC showed delayed mild reaction, a response of ischemic EC was immediate and intensifies with time. In normal EC there was a depletion of GSH from 24.1±1.5 to 18.5±1.1 nM/mg protein (p<0.05) at 18 hr; however without changes in TBARS and only insignificant expression of VACAM-1 in the absence of apoptosis. On the contrary, CM showed to strongly affect ischemic EC causing further redox imbalance. At 2 hr GSH dropped from 13.6±0.9 to 10.3±0.6 (p<0.05) and progressively decreased to 9.1±0.5 (p<0.01) at 4 hr and 7.5±0.4 (p<0.001) at 18 hr. TBARS progressively increased from 0.84±0.03 to 0.98±0.04 nM/mg protein (p<0.05) at 2 hr, 1.1±0.07 (p<0.01) at 4 hr and 1.44±0.12 (p<0.001) at 18 hr. A slight expression of VACAM-1 was noted at 4 hr that reached the peak at 18 hr (p<0.01). Early apoptosis was detected at 18 hr. HemoTech treatment effectively restored the redox balance in ischemic EC and alleviated inflammatory and apoptotic responses to CM. All tested parameters were in normal range. Conclusions: Low-osmolar nonionic iodinated CM is well tolerated by normal but not ischemic coronary artery EC. HemoTech blood substitute with pharmacological properties of ATP, adenosine and GSH protects ischemic EC against CM-induced toxicity.

The Future of Lymphedema: Potential Therapeutic Targets for Treatment.

This review aims to summarize the current knowledge regarding the pharmacological interventions studied in both experimental and clinical trials for secondary lymphedema. Lymphedema is a progressive disease that results in tissue swelling, pain, and functional disability. The most common cause of secondary lymphedema in developed countries is an iatrogenic injury to the lymphatic system during cancer treatment. Despite its high incidence and severe sequelae, lymphedema is usually treated with palliative options such as compression and physical therapy. However, recent studies on the pathophysiology of lymphedema have explored pharmacological treatments in preclinical and early phase clinical trials. Many potential treatment options for lymphedema have been explored throughout the past two decades including systemic agents and topical approaches to decrease the potential toxicity of systemic treatment. Treatment strategies including lymphangiogenic factors, anti-inflammatory agents, and anti-fibrotic therapies may be used independently or in conjunction with surgical approaches.

Amyloid accumulation and functional connectivity in normal aging and preclinical Alzheimer’s Disease: a longitudinal analysis of the OASIS‐3 cohort

AbstractBackgroundPathological changes in the brain begin accumulating decades before the appearance of cognitive symptoms in Alzheimer’s Disease (AD). The deposition of amyloid‐beta (Aβ) proteins and other neurotoxic changes occur, leading to disruption in functional connections between brain networks. Discrete characterization of the changes that take place in preclinical AD has the potential to help treatment development by targeting the neuropathological mechanisms to prevent cognitive decline and dementia from occurring entirely. Previous research has focused on cross sectional differences in the brains of patients with mild cognitive impairment (MCI) or AD and healthy controls or has concentrated on the stages immediately preceding cognitive symptoms. The present study emphasizes early preclinical phases of neurodegeneration.MethodWe use a longitudinal approach to examine the brain changes that take place during early stages of cognitive decline in the OASIS‐3 dataset. Among 1098 participants, 274 passed the inclusion criteria (i.e. had at least two repeated measures of cognitive status and amyloid levels). We use mixed‐effect linear models to examine rates of amyloid accumulation in those who showed cognitive decline compared to those who did not change in cognitive status.ResultOver 90% of participants were healthy at baseline. Over 8‐10 years, some participants progressed to very mild dementia (n = 48) while others stayed healthy (n = 226). Participants with cognitive decline have faster amyloid accumulation and greater increases in functional connectivity in similar brain regions, namely among lateral temporal, motor, and some lateral prefrontal cortex areas. These changes were linked to increases in functional connectivity of select default mode, frontoparietal and motor components.ConclusionOur findings advance the understanding of initial amyloid staging in preclinical Alzheimer’s, as well as contribute to the determination of the functional changes that occur among brain network components as the disease progresses.