Therapeutic Screening Research Articles

Repurposing FDA-Approved Anastrozole-based Drugs for Breast Cancer through Drug-Drug Transcriptomic Similarity and Cavity Detection Guided Blind Docking

Breast cancer remains a significant global health concern, necessitating innovative strategies for drug discovery and development. Repurposing existing drugs offers a promising avenue, leveraging the wealth of information available on food and drug administration (FDA) approved compounds. This study investigates the repurposing potential of anastrozole-based drugs, already established for their efficacy in certain conditions, for breast cancer treatment. This research presents a multifaceted investigation into the identification, structural refinement, and virtual screening of potential therapeutics targeting the aromatase CYP19A enzyme, a crucial player in hormone-related conditions such as breast cancer. The study commences with the identification of anastrozole-based drugs exhibiting transcriptomic profiles closely resembling known breast cancer therapeutics. Through a comprehensive analysis, a subset of compounds demonstrates high transcriptomic similarity, suggesting shared molecular pathways and target interactions. Notably, anastrozole, a well-known aromatase inhibitor, emerges as a top candidate, highlighting its potential in breast cancer treatment. The crystallographic structure of aromatase CYP19A is subjected to meticulous preprocessing using the PDB-REDO server, resulting in significant improvements in various validation metrics. Structural changes, including alterations in rotamers, removal of water molecules, and peptide flips, indicate the success of the refinement process in enhancing the accuracy of the protein model. The refined structure serves as a reliable foundation for subsequent studies. Further, structure-based cavity detection unveils potential binding sites on the aromatase enzyme. Docking studies employing the cb-dock server elucidate the interaction patterns and binding affinities of selected compounds within these cavities. Anastrozole, along with other candidates like dolasetron and stiripentol, exhibits promising binding scores and interacts with specific residues crucial for enzyme activity. This integrative approach, combining transcriptomic similarity analysis, structural refinement, and virtual screening, provides valuable insights into potential lead compounds for the inhibition of aromatase in breast cancer therapy. The identified compounds offer a starting point for further experimental validation and drug development. The most promising compound that can be repurposed for as an aromatase inhibitor is dolasetron. Overall, this research contributes to the ongoing efforts to leverage computational methodologies for the rational design of targeted therapeutics against hormone-related disorders.

IMPLICATIONS OF RESIDENT AND ENVIRONMENTAL CHARACTERISTICS ON RESIDENT-TO-RESIDENT AGGRESSION

Abstract This study examines factors associated with resident-to-resident aggression (RRA) in long term care facilities in Hong Kong. A total of 800 personal care worker participated. Participants averaged 42.03 years of age (SD=7.63), were mostly female (92.7%), married (79.1%) and reported an average of 6.28 years of experience in long term care. 96.9 percent of the participants provided care to residents with dementia but 58.9% considered the training they received insufficient. RRA was common: All participants reported having witnessed verbal aggression (100%), 18% disruptive behaviors, 11.8% physical violence, and 3.1% sexual aggression. The following analysis used a subset of 412 participant from 29 long term care facilities where environmental assessments were conducted. General linear model analysis showed that RRA was associated with both resident and environmental characteristics. Victims’ neuropsychiatry symptoms, perpetrator male gender and neuropsychiatric symptoms were associated with higher RRA. Separation of residents with and without dementia, inappropriate use of nursing unit for residents as measured by Therapeutic Environment Screening Survey for Nursing Homes (TESS-NH), lack of functional and attractiveness courtyard, less cleanliness of spaces, insufficient lighting and insufficient orientation / cueing as measured by Professional Environmental Assessment Procedure (PEAP) were also associated with higher RRA. There is an urgent need to prevent and intervene RRA in long term care facilities. Improving management of neuropsychiatric symptoms and creating a supportive and accommodating environment could be helpful in this regard.

A resource of induced pluripotent stem cell (iPSC) lines including clinical, genomic, and cellular data from genetically isolated families with mood and psychotic disorders

Genome-wide (GWAS) and copy number variant (CNV) association studies have reproducibly identified numerous risk alleles associated with bipolar disorder (BD), major depressive disorder (MDD), and schizophrenia (SCZ), but biological characterization of these alleles lags gene discovery, owing to the inaccessibility of live human brain cells and inadequate animal models for human psychiatric conditions. Human-derived induced pluripotent stem cells (iPSCs) provide a renewable cellular reagent that can be differentiated into living, disease-relevant cells and 3D brain organoids carrying the full complement of genetic variants present in the donor germline. Experimental studies of iPSC-derived cells allow functional characterization of risk alleles, establishment of causal relationships between genes and neurobiology, and screening for novel therapeutics. Here we report the creation and availability of an iPSC resource comprising clinical, genomic, and cellular data obtained from genetically isolated families with BD and related conditions. Results from the first 324 study participants, 61 of whom have validated pluripotent clones, show enrichment of rare single nucleotide variants and CNVs overlapping many known risk genes and pathogenic CNVs. This growing iPSC resource is available to scientists pursuing functional genomic studies of BD and related conditions.

Human retinal organoids with an OPA1 mutation are defective in retinal ganglion cell differentiation and function

Autosomal dominant optic atrophy (ADOA), mostly caused by heterozygous OPA1 mutations and characterized by retinal ganglion cell (RGC) loss and optic nerve degeneration, is one of the most common types of inherited optic neuropathies. Previous work using a two-dimensional (2D) differentiation model of induced pluripotent stem cells (iPSCs) has investigated ADOA pathogenesis but failed to agree on the effect of OPA1 mutations on RGC differentiation. Here, we use 3D retinal organoids capable of mimicking invivo retinal development to resolve the issue. We generated isogenic iPSCs carrying the hotspot OPA1 c.2708_2711delTTAG mutation and found that the mutant variant caused defective initial and terminal differentiation and abnormal electrophysiological properties of organoid-derived RGCs. Moreover, this variant inhibits progenitor proliferation and results in mitochondrial dysfunction. These data demonstrate that retinal organoids coupled with gene editing serve as a powerful tool to definitively identify disease-related phenotypes and provide valuable resources to further investigate ADOA pathogenesis and screen for ADOA therapeutics.

Ex vivo precision-cut liver slices model disease phenotype and monitor therapeutic response for liver monogenic diseases

Background In academic research and the pharmaceutical industry, in vitro cell lines and in vivo animal models are considered as gold standards in modelling diseases and assessing therapeutic efficacy. However, both models have intrinsic limitations, whilst the use of precision-cut tissue slices can bridge the gap between these mainstream models. Precision-cut tissue slices combine the advantage of high reproducibility, studying all cell sub-types whilst preserving the tissue matrix and extracellular architecture, thereby closely mimicking a mini-organ. This approach can be used to replicate the biological phenotype of liver monogenic diseases using mouse models. Methods Here, we describe an optimised and easy-to-implement protocol for the culture of sections from mouse livers, enabling its use as a reliable ex-vivo model to assess the therapeutic screening of inherited metabolic diseases Results We show that precision-cut liver sections can be a reliable model for recapitulating the biological phenotype of inherited metabolic diseases, exemplified by common urea cycle defects such as citrullinemia type 1 and argininosuccinic aciduria, caused by argininosuccinic synthase (ASS1) and argininosuccinic lyase (ASL) deficiencies respectively. Conclusions Therapeutic response to gene therapy such as messenger RNA replacement delivered via lipid nanoparticles can be monitored, demonstrating that precision-cut liver sections can be used as a preclinical screening tool to assess therapeutic response and toxicity in monogenic liver diseases.

Advances in high throughput cell culture technologies for therapeutic screening and biological discovery applications.

Cellular phenotypes and functional responses are modulated by the signals present in their microenvironment, including extracellular matrix (ECM) proteins, tissue mechanical properties, soluble signals and nutrients, and cell-cell interactions. To better recapitulate and analyze these complex signals within the framework of more physiologically relevant culture models, high throughput culture platforms can be transformative. High throughput methodologies enable scientists to extract increasingly robust and broad datasets from individual experiments, screen large numbers of conditions for potential hits, better qualify and predict responses for preclinical applications, and reduce reliance on animal studies. High throughput cell culture systems require uniformity, assay miniaturization, specific target identification, and process simplification. In this review, we detail the various techniques that researchers have used to face these challenges and explore cellular responses in a high throughput manner. We highlight several common approaches including two-dimensional multiwell microplates, microarrays, and microfluidic cell culture systems as well as unencapsulated and encapsulated three-dimensional high throughput cell culture systems, featuring multiwell microplates, micromolds, microwells, microarrays, granular hydrogels, and cell-encapsulated microgels. We also discuss current applications of these high throughput technologies, namely stem cell sourcing, drug discovery and predictive toxicology, and personalized medicine, along with emerging opportunities and future impact areas.

Developing Topics.

Goal 1 of the National Plan to Address Alzheimer's Disease (AD) is to prevent and effectively treat AD and AD-related dementias (ADRD) by 2025. Toward this goal, NINDS plans 14 ADRD funding opportunities for 2024 to support training, basic mechanisms, translational, and clinical research for ADRD. Here, we describe ADRD translational therapy development initiatives, which include 1) multi-target validation, 2) early-stage therapy development, 3) genome editing and 4) animal model development and validation. Note, ADRD specific funding opportunities are not required. ADRD researchers can also take advantage of NINDS existing programs such as the NIH Blueprint for Neurotherapeutics (BPN) for Small Molecule or Biologic therapy development. These programs support optimization, manufacture, IND-enabling studies and first in human clinical trials through a combination of grants and complementary access to NIH-provided CROs and consultants. All concepts discussed here are approved, however NIH cannot guarantee future funding opportunities, award mechanisms, or funding allocation. A funding opportunity is only official when published in the NIH Guide. Multi-Target Validation for ADRD concept has the goal to support research that would validate multiple targets for future therapy development that better reflect the complexity of disease. Early-Stage Therapy Development concept would support the development of small molecule or biologic therapeutics and would be a reissue of RFA-NS-22-059, which supported assay development, therapeutic screening, optimization, and in vivo efficacy work in preparation for later-stage programs. Genome Editing for ADRD concept would support early translational research focused on somatic cell gene editing with the expectation that studies will address feasibility of the gene editing approach and would be a reissue of RFA-NS-23-017. PAR-23-154 "Development and Validation of Models for Alzheimer's Disease-Related Dementias (ADRD)" supports the development and validation of clinically- and pathophysiologically-relevant models of ADRD that can be used for future therapy development. Models should reflect multiple aspects of the human condition and include internal, face, construct, and predictive validation to the extent possible. Through these initiatives, NINDS hopes to provide a comprehensive pipeline for ADRD therapy development from target validation to the clinic.

A human stem cell model of development and pathology of basal forebrain cholinergic neurons

AbstractBackgroundBasal forebrain cholinergic neurons (BFCNs) in the nucleus basalis regulate attention, memory, learning and processing of information related to cognitive function throughout the lifespan. Degeneration of BFCNs is strongly correlated with cognitive function and degeneration of the BFCN projection system is an early event in Alzheimer’s disease (AD) pathogenesis and in Down syndrome (DS, trisomy 21) that is characterized by intellectual impairment at birth and AD pathology in middle age. Understanding the vulnerability of BFCN neurons will help us understand the underlying mechanisms of neurodegeneration in both AD and DS.MethodWe generated BFCNs from trisomy 21 induced pluripotent stem cells (iPSCs) and isogenic controls through addition of morphogens and growth factors following developmental principles. The efficiency of the protocol was assessed through immunofluorescence for choline acetyl transferase (ChAT).ResultWe established a protocol to generate BFCNs from control and trisomy 21 iPSCs through a stepwise process of 1) ventralization of neural progenitors with SHH or SHH agonist, 2) regulation of LHX8 expression with NGF and, 3) regulation of ISLET1 expression and BFCN survival with BMP9. The protocol generated approximately 60% BFCNs from both trisomy 21 and control iPSCs.ConclusionFew differentiation protocols have been established to derive BFCNs from iPSCs. Our iPSC model enables analysis of pathological markers, identification of disrupted molecular pathways, manipulation of conditions to affect neurodegeneration and screening of potential therapeutics in human BFCNs.

P182: Effects of Physical Environment on Quality of Life of Residents in Dementia Facilities in Canada & South Korea: A Longitudinal Observational Study

Approximately 44% of new residents of care facilities in Korea were diagnosed with dementia (Song, Park & Kim, 2013), and in Canada, about one-third of older adults younger than 80 who have been diagnosed with dementia live in long-term care facilities (Canadian Institute for Health Information, 2018). Due to the rapid increase of these figures in the future, continuing to provide assistance services and appropriate environment for residents with dementia could be challenging for both countries.This longitudinal observational study aims to examine whether residents with dementia in long-term care facilities with variability in physical environment attributions in Vancouver (N=11), Canada and Seoul (N=9), South Korea had a distinction in their quality of life (QoL). Physical environmental assessment was conducted using the Therapeutic Environment Screening Survey for Nursing Homes (TESS-NH) (Sloane et al., 2002). QoL was assessed three times over one year using Dementia Care Mapping tool (DCM) (University of Bradford, 2010). The results of the study demonstrated that the residents with dementia living in an institutional large-scale setting showed statistically more withdrawn behavior and spent more time to be negative mood or affect compared to the ones in a small-scale setting. This study also found that the number of potential positive behaviors of residents in a small-scale setting was three times higher than that of residents in an institutional large-scale setting. When looking at the distinction between two countries in the behavior category with a large average time difference, the residents with dementia in Korea had shorter meal/dessert times compared to those in Canada. The study supports that the small-scale homelike environment is intensely associated with a therapeutic environment for older adults with dementia.

A Zebrafish Model of Parn-Deficiency Reveals Conserved Mechanisms Underlying Marrow Failure and Opportunities for Therapeutic Amelioration of Hematopoiesis

Dyskeratosis congenita (DC) is a rare inherited bone marrow failure syndrome (IBMFS) and a telomere biology disorder that is characterized by physical malformations such as nail dystrophy, oral leukoplakia, reticular hyperpigmentation, multilineage cytopenia and immunodeficiency. Definitive treatment for the hematological complications is hematopoietic stem cell transplantation (HSCT), which is associated with high risk of organ toxicity and engraftment failure in this population. Further, DC patients are prone to developing myelodysplastic syndrome (MDS) and solid tumors. Germline mutations in the poly(A)-specific ribonuclease (PARN) gene are associated with Hoyeraal-Hreidarsson syndrome, the most severe form of DC. PARN encodes a deadenylase enzyme involved in the 3'-end processing of various RNAs including the telomerase RNA component (TERC). The clinical heterogeneity of phenotypes and the paucity of primary human samples necessitate the use of animal models such as zebrafish ( Danio rerio), given their highly conserved genetic program, to elucidate underlying biology. Zebrafish have a single PARN ortholog with 64% sequence identity at the amino acid level. Using CRISPR-Cas9 mutagenesis, we generated a 1.3 kb deletion in the zebrafish parn gene, predicted to result in premature protein truncation. To measure the hematopoietic output of the parn mutants, we performed whole-mount in situ hybridization (WISH) for key hematopoietic lineage markers. At 48 hours post-fertilization (hpf), we found a reduction in lcp1 + total leukocytes, mpx + neutrophils and hbbe3 + mature erythrocytes. Impaired erythroid differentiation was accompanied by an accumulation of gata1 + erythroid progenitors. This finding was further corroborated by o-dianisidine staining, showing reduced hemoglobinized erythrocytes. Thus, erythrocyte and leukocyte differentiation are compromised in our zebrafish parn mutant, phenocopying the multilineage cytopenia observed in DC patients. Flow cytometry of the whole kidney marrow (WKM, human bone marrow equivalent) at 12 months of age revealed an increase in hematopoietic stem and progenitor cells in the parn mutants. Giemsa staining of WKM touchpreps further identified an increase in myeloid-biased progenitors. Next, we investigated the consequences of Parn loss on telomerase activity. Using the telomere repeat amplification protocol, we found reduced telomerase activity in the WKMs of parn mutant zebrafish. Using 3' rapid amplification of cDNA ends, we observed longer ends in the terc amplicons of the parn mutants, reminiscent of defective deadenylation of human TERC RNA in the setting of PARN mutations. Exogenous thymidine supplementation has been shown to support telomere elongation in human cells. Following treatment of zebrafish embryos with thymidine nucleoside, we observed an improvement in erythrocyte numbers in the parn mutants by o-dianisidine staining at 48 hpf. Collectively, our preliminary findings suggest that telomerase-dependent mechanisms may contribute to the erythropenia seen in parn-mutant zebrafish. In summary, we generated a robust zebrafish model carrying a germline parn mutation that phenocopies the molecular and hematologic features seen in DC patients. Our model holds promise as a new in vivo tool for investigating DC-related disease mechanisms and as a platform for preclinical therapeutic screening.

Multi-level characteristics recognition of cancer core therapeutic targets and drug screening for a broader patient population

Multi-level characteristics recognition of cancer core therapeutic targets and drug screening for a broader patient population

A High-Throughput, High-Containment Human Primary Epithelial Airway Organ-on-Chip Platform for SARS-CoV-2 Therapeutic Screening.

COVID-19 emerged as a worldwide pandemic in early 2020, and while the rapid development of safe and efficacious vaccines stands as an extraordinary achievement, the identification of effective therapeutics has been less successful. This process has been limited in part by a lack of human-relevant preclinical models compatible with therapeutic screening on the native virus, which requires a high-containment environment. Here, we report SARS-CoV-2 infection and robust viral replication in PREDICT96-ALI, a high-throughput, human primary cell-based organ-on-chip platform. We evaluate unique infection kinetic profiles across lung tissue from three human donors by immunofluorescence, RT-qPCR, and plaque assays over a 6-day infection period. Enabled by the 96 devices/plate throughput of PREDICT96-ALI, we also investigate the efficacy of Remdesivir and MPro61 in a proof-of-concept antiviral study. Both compounds exhibit an antiviral effect against SARS-CoV-2 in the platform. This demonstration of SARS-CoV-2 infection and antiviral dosing in a high-throughput organ-on-chip platform presents a critical capability for disease modeling and therapeutic screening applications in a human physiology-relevant in vitro system.

Clinical study of ferredoxin-reductase-related mitochondriopathy: Genotype-phenotype correlation and proposal of ancestry-based carrier screening in the Mexican population

PurposeFerredoxin reductase (FDXR) is a flavoprotein that functions in both iron sulfur cluster biogenesis and steroid biosynthesis pathways in the mitochondria. Not surprisingly, loss of FDXR function causes severe mitochondrial diseases in humans. Although several FDXR-related mitochondriopathy (FRM) cohorts have been reported in the literature, further characterization of the natural history of FRM is warranted. MethodsTo better understand the spectrum of FRM, a natural history study of FRM was performed. New cases were added to previously reported FRM cases for analysis (n = 62 cases). ResultsOptic atrophy, movement disorder, and developmental delay were frequent findings. Mortality is high, with 18% of patients, often infants, passing from complications. Notably, 25% of cases were homozygous or compound heterozygous for the previously reported p.Arg386Trp “hotspot” variant. Of the obtained ancestry, all but 1 individual heterozygous for the p.Arg386Trp variant was Hispanic, with many reporting Mexican heritage. Utilizing recent large-scale genome sequencing surveys, the carrier frequency of the p.Arg386Trp variant was estimated as 1 of 185 in the Mexican population. ConclusionGiven the high mortality of FRM and carrier frequency of the common variant, consideration of a new approach for population carrier screening and development of therapeutics for affected individuals is needed.

174P Unlocking the potential of blood-based biomarkers in pancreatic cancer for early detection and therapeutic screening

174P Unlocking the potential of blood-based biomarkers in pancreatic cancer for early detection and therapeutic screening

Urine-derived stem cells in neurological diseases: current state-of-the-art and future directions.

Stem cells have potential applications in the field of neurological diseases, as they allow for the development of new biological models. These models can improve our understanding of the underlying pathologies and facilitate the screening of new therapeutics in the context of precision medicine. Stem cells have also been applied in clinical tests to repair tissues and improve functional recovery. Nevertheless, although promising, commonly used stem cells display some limitations that curb the scope of their applications, such as the difficulty of obtention. In that regard, urine-derived cells can be reprogrammed into induced pluripotent stem cells (iPSCs). However, their obtaining can be challenging due to the low yield and complexity of the multi-phased and typically expensive differentiation protocols. As an alternative, urine-derived stem cells (UDSCs), included within the population of urine-derived cells, present a mesenchymal-like phenotype and have shown promising properties for similar purposes. Importantly, UDSCs have been differentiated into neuronal-like cells, auspicious for disease modeling, while overcoming some of the shortcomings presented by other stem cells for these purposes. Thus, this review assesses the current state and future perspectives regarding the potential of UDSCs in the ambit of neurological diseases, both for disease modeling and therapeutic applications.

Risk model-based management for second primary lung cancer among lung cancer survivors through a validated risk prediction model.

Recent therapeutic advances and screening technologies have improved survival among patients with lung cancer, who are now at high risk of developing second primary lung cancer (SPLC). Recently, an SPLC risk-prediction model (called SPLC-RAT) was developed and validated using data from population-based epidemiological cohorts and clinical trials, but real-world validation has been lacking. The predictive performance of SPLC-RAT was evaluated in a hospital-based cohort of lung cancer survivors. The authors analyzed data from 8448 ever-smoking patients diagnosed with initial primary lung cancer (IPLC) in 1997-2006 at Mayo Clinic, with each patient followed for SPLC through 2018. The predictive performance of SPLC-RAT and further explored the potential of improving SPLC detection through risk model-based surveillance using SPLC-RAT versus existing clinical surveillance guidelines. Of 8448 IPLC patients, 483 (5.7%) developed SPLC over 26,470 person-years. The application of SPLC-RAT showed high discrimination area under the receiver operating characteristics curve: 0.81). When the cohort was stratified by a 10-year risk threshold of ≥5.6% (i.e., 80th percentile from the SPLC-RAT development cohort), the observed SPLC incidence was significantly elevated in the high-risk versus low-risk subgroup (13.1% vs. 1.1%, p<1×10-6 ). The risk-based surveillance through SPLC-RAT (≥5.6% threshold) outperformed the National Comprehensive Cancer Network guidelines with higher sensitivity (86.4% vs. 79.4%) and specificity (38.9% vs. 30.4%) and required 20% fewer computed tomography follow-ups needed to detect one SPLC (162 vs. 202). In a large, hospital-based cohort, the authors validated the predictive performance of SPLC-RAT in identifying high-risk survivors of SPLC and showed its potential to improve SPLC detection through risk-based surveillance. Lung cancer survivors have a high risk of developing second primary lung cancer (SPLC). However, no evidence-based guidelines for SPLC surveillance are available for lung cancer survivors. Recently, an SPLC risk-prediction model was developed and validated using data from population-based epidemiological cohorts and clinical trials, but real-world validation has been lacking. Using a large, real-world cohort of lung cancer survivors, we showed the high predictive accuracy and risk-stratification ability of the SPLC risk-prediction model. Furthermore, we demonstrated the potential to enhance efficiency in detecting SPLC using risk model-based surveillance strategies compared to the existing consensus-based clinical guidelines, including the National Comprehensive Cancer Network.

Evaluation of Measurement Properties of Supportive Environment Assessment Scales for Dementia Special Care Units: A COSMIN Systematic Review

ObjectivesTo identify, critically appraise, compare, and summarize the measurement properties of existing instruments that assess the supportive environment of dementia special care units (DSCUs). DesignSystematic review of measurement properties consistent with Consensus-based standards for the selection of health measurement instruments (COSMIN) guidelines. Settings and ParticipantsPubMed, Embase, Web of Science, CINAHL, CNKI, Wanfang, VIP, and SinoMed were searched from inception to July 21, 2023. Studies that (1) measured the supportive environment for DSCUs using any type of assessment instrument and (2) evaluated 1 or more psychometric properties of a DSCU's supportive environment assessment instruments were included. MethodsTwo reviewers independently screened, selected, extracted data, and assessed risk of bias. ResultsFourteen studies were identified that reported the psychometric properties of 8 assessment instruments. The Therapeutic Environment Screening Survey for Nursing Homes (TESS-NH) exhibited relatively better results on methodological risk of bias and quality of the psychometric properties. None of the instruments reported the evaluations on hypothesis testing, cross-cultural validity/measurement invariance, measurement error, or responsiveness. Based on the summary of 32 dimensions from 8 assessment instruments, this review established 7 functional constructs for the supportive environment for DSCUs: safety maintenance, space design, external resources, sensory stimulation, humanistic care, residual function development, and professional care. In addition, this study also initially developed a conceptual framework for the supportive environment of DSCUs. Conclusion and ImplicationsTESS-NH received the rating of “best methodological quality” and outperformed other weakly recommended scales. Further studies should pay attention to developing or revalidating scales for assessing the supportive environment of DSCUs in large multicenter samples following the COSMIN methodology. Furthermore, the conceptual framework for the DSCU supportive environment will provide a theoretical reference for facilitating their hierarchical establishment and governance within diverse long-term care facilities by state authorities.

Leukocyte immunoglobulin-like receptor B2 overexpression as a promising therapeutic target and noninvasive screening biomarker for colorectal cancer.

Colorectal cancer (CRC) has become the second most deadly malignancy in the world, and the exploration of screening markers and precise therapeutic targets is urgent. Our previous research identified leukocyte immunoglobulin-like receptor B2 (LILRB2) protein as a characteristic protein of CRC, but the association between LILRB2 expression and clinicopathological features, the internal mechanism related to CRC progression, and screening diagnostic efficacy are not clear. Therefore, we hypothesized that LILRB2 is significantly highly expressed in CRC tissues, correlated with advanced stage and a poor prognosis, and could be used as a therapeutic target and potential screening biomarker for CRC. To explore whether LILRB2 can be used as a potential therapeutic target and noninvasive screening biomarker for CRC. Patients who underwent radical surgery for CRC at China-Japan Friendship Hospital between February 2021 and October 2022 were included. Cancer and paracancerous tissues were collected to verify LILRB2 expression, and the association between LILRB2 expression and clinicopathological features was analysed. Serum was collected from CRC patients, adenoma patients and healthy controls during the same period to assess the diagnostic value of LILRB2 as a noninvasive screening biomarker, and its diagnostic value was further compared with that of the traditional markers carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). A total of 58 CRC patients were included, and LILRB2 protein was significantly overexpressed in cancer tissues compared with paracancerous tissues (P < 0.001). Angiopoietin-like protein 2 (ANGPTL2) protein, as the ligand of LILRB2, was synergistically overexpressed in CRC tissues (P < 0.001), and overexpression of LILRB2 and ANGPTL2 protein was significantly correlated with poor to moderate differentiation, vascular involvement, lymph node metastasis, distant metastasis, advanced tumor-node-metastasis stage and a poor prognosis (P < 0.05), which suggested that LILRB2 and ANGPTL2 are closely associated with CRC progression. In addition, serum LILRB2 concentrations increased stepwise in healthy individuals, adenoma patients and CRC patients with statistically significant differences. The sensitivity of serum LILRB2 for the diagnosis of CRC was 89.74%, the specificity was 88.89%, the area under the curve was 0.95, and the diagnostic efficacy was better than that of conventional CEA and CA19-9. LILRB2 protein can be used as a potential novel therapeutic target and noninvasive screening biomarker for CRC, which is beneficial for early screening and precise treatment.

A sticky-end probe biosensor for homogeneous detection of transcription factor binding activity.

Transcription factors are essential regulators of various physiological and pathological processes. However, detecting transcription factor-DNA binding activities is often time-consuming and labor-intensive. Homogeneous biosensors that are compatible with mix-and-measure protocols have the potential to simplify the workflow for therapeutic screening and disease diagnostics. In this study, we apply a combined computational-experimental approach to investigate the design of a sticky-end probe biosensor, where the transcription factor-DNA complex stabilizes the fluorescence resonance energy transfer signal of the donor-acceptor pair. We design a sticky-end biosensor for the SOX9 transcription factor based on the consensus sequence and characterize its sensing performance. A systems biology model is also developed to investigate the reaction kinetics and optimize the operating conditions. Taken together, our study provides a conceptual framework for the design and optimization of sticky-end probe biosensors for homogeneous detection of transcription factor-DNA binding activity.

World's First Long-Term Colorectal Cancer Model by 3D Bioprinting as a Mechanism for Screening Oncolytic Viruses.

Long-term modelization of cancer as it changes in the human body is a difficult goal, particularly when designing and testing new therapeutic strategies. This becomes even more difficult with metastasis modeling to show chemotherapeutic molecule delivery directly to tumoral cells. Advanced therapeutics, including oncolytic viruses, antibody-based and cell-based therapies are increasing. The question is, are screening tests also evolving? Next-generation therapeutics need equally advanced screening tests, which whilst difficult to achieve, are the goal of our work here, creating models of micro- and macrotumors using 3D bioprinting. We developed advanced colorectal cancer tumor processing techniques to provide options for cellular expansion, microtumor printing, and long-term models, which allow for the evaluation of the kinetics of penetration testing, therapeutic success, targeted therapies, and personalized medicine. We describe how we tested tumors from a primary colorectal patient and, applying 3D bioprinting, matured long-term models for oncolytic metastatic screening. Three-dimensional microtumors were kept alive for the longest time ever recorded in vitro, allowing longitudinal studies, screening of oncolytic viruses and realistic modelization of colorectal cancer. These 3D bioprinted models were maintained for around 6 months and were able to demonstrate the effective delivery of a product to the tumoral environment and represent a step forward in therapeutic screening.