Reverse Translational Approach Research Articles

Correlation between tooth decay and insulin resistance in normal weight males prompts a role for myo-inositol as a regenerative factor in dentistry and oral surgery: a feasibility study.

In an era of precision and stratified medicine, homogeneity in population-based cohorts, stringent causative entry, and pattern analysis of datasets are key elements to investigate medical treatments. Adhering to these principles, we collected in vivo and in vitro data pointing to an insulin-sensitizing/insulin-mimetic effect of myo-inositol (MYO) relevant to cell regeneration in dentistry and oral surgery. Confirmation of this possibility was obtained by in silico analysis of the relation between in vivo and in vitro results (the so-called bed-to-benchside reverse translational approach). Fourteen subjects over the 266 screened were young adult, normal weight, euglycemic, sedentary males having normal appetite, free diet, with a regular three-times-a-day eating schedule, standard dental hygiene, and negligible malocclusion/enamel defects. Occlusal caries were detected by fluorescence videoscanning, whereas body composition and energy balance were estimated with plicometry, predictive equations, and handgrip. Statistically significant correlations (Pearson r coefficient) were found between the number of occlusal caries and anthropometric indexes predicting insulin resistance (IR) in relation to the abdominal/visceral fat mass, fat-free mass, muscular strength, and energy expenditure adjusted to the fat and muscle stores. This indicated a role for IR in affecting dentin reparative processes. Consistently, in vitro administration of MYO to HUVEC and Swiss NIH3T3 cells in concentrations corresponding to those administered in vivo to reduce IR resulted in statistically significant cell replication (ANOVA/Turkey tests), suggesting that MYO has the potential to counteract inhibitory effects of IR on dental vascular and stromal cells turnover. Finally, in in silico experiments, quantitative evaluation (WOE and information value) of a bioinformatic Clinical Outcome Pathway confirmed that in vitro trophic effects of MYO could be transferred in vivo with high predictability, providing robust credence of its efficacy for oral health. Our reverse bed-to-benchside data indicate that MYO might antagonize the detrimental effects of IR on tooth decay. This provides feasibility for clinical studies on MYO as a regenerative factor in dentistry and oral surgery, including dysmetabolic/aging conditions, bone reconstruction in oral destructive/necrotic disorders, dental implants, and for empowering the efficacy of a number of tissue engineering methodologies in dentistry and oral surgery.

Anti-inflammatory Effects of Bacteroidota Strains Derived From Outstanding Donors of Fecal Microbiota Transplantation for the Treatment of Ulcerative Colitis.

The proportion of certain Bacteroidota species decreased in patients with ulcerative colitis, and the recovery of Bacteroidota is associated with the efficacy of fecal microbiota transplantation therapy. We hypothesized that certain Bacteroidota may advance ulcerative colitis treatment. Accordingly, we aimed to evaluate the anti-inflammatory effects of Bacteroidota strains isolated from donors. Donors with proven efficacy of fecal microbiota transplantation for ulcerative colitis were selected, and Bacteroidota strains were isolated from their stools. The immune function of Bacteroidota isolates was evaluated through in vitro and in vivo studies. Twenty-four Bacteroidota strains were isolated and identified. Using an in vitro interleukin (IL)-10 induction assay, we identified 4 Bacteroidota strains with remarkable IL-10-induction activity. Of these, an Alistipes putredinis strain exhibited anti-inflammatory effects in a mouse model of colitis induced by sodium dextran sulfate and oxazolone. However, 16S rRNA gene-based sequencing analysis of A. putredinis cultures in the in vivo study revealed unexpected Veillonella strain contamination. A second in vitro study confirmed that the coculture exhibited an even more potent IL-10-inducing activity. Furthermore, the production of A. putredinis-induced IL-10 was likely mediated via toll-like receptor 2 signaling. This study demonstrated that A. putredinis, a representative Bacteroidota species, exhibits anti-inflammatory effects in vivo and in vitro; however, the effects of other Bacteroidota species remain unexplored. Our fecal microbiota transplantation-based reverse translation approach using promising bacterial species may represent a breakthrough in microbiome drug development for controlling dysbiosis during ulcerative colitis.

Advancing clinical development for neuronopathic Hunter syndrome through a quantitatively-driven reverse translation framework.

A quantitatively-driven evaluation of existing clinical data and associated knowledge to accelerate drug discovery and development is a highly valuable approach across therapeutic areas, but remains underutilized. This is especially the case for rare diseases for which development is particularly challenging. The current work outlines an organizational framework to support a quantitatively-based reverse translation approach to clinical development. This approach was applied to characterize predictors of the trajectory of cognition in Hunter syndrome (Mucopolysaccharidosis Type II; MPS-II), a rare X-linked lysosomal storage disorder, highly heterogeneous in its course. Specifically, we considered ways to refine target populations based on age, cognitive status, and biomarkers, that is, cerebrospinal fluid glycosaminoglycans (GAG), at trial entry. Data from a total of 138 subjects (age range 2.5 to 10.1 years) from Takeda-sponsored internal studies and external natural history studies in MPS-II were included. Quantitative analyses using mixed-effects models were performed to characterize the relationships between neurocognitive outcomes and potential indicators of disease progression. Results revealed a specific trajectory in cognitive development across age with an initial progressive phase, followed by a plateau between 4 and 8 years and then a variable declining phase. Additionally, results suggest a faster decline in cognition among subjects with lower cognitive scores or with higher cerebrospinal fluid GAG at enrollment. These results support differences in the neurocognitive course of MPS-II between distinct groups of patients based on age, cognitive function, and biomarker status at enrollment. These differences should be considered when designing future clinical trials.

Inactivation of phosphodiesterase-4B gene in rat nucleus accumbens shell by CRISPR/Cas9 or positive allosteric modulation of the protein affects the motivation to chronically self-administer nicotine

Large scale human genome wide association studies (GWAS) have identified a growing pool of genes associated with cigarette smoking. One of the most prominent, phosphodiesterase-4B (PDE4B), has been associated with multiple smoking phenotypes. Although PDE4B modulates the half-life of neuronal cAMP, its precise role in smoking behaviors is unknown. To address this knowledge gap, we used a reverse translational approach. We inactivated PDE4B in bilateral medial nucleus accumbens shell (NAcs) neurons by injecting AAV containing a specific gRNA in female transgenic Cas9+ Long Evans rats. These rats then were given 23-h chronic access to nicotine intravenous self-administration (IVSA) under a schedule of increasing fixed ratios (FR). With the increased effort required at FR7, nicotine SA (i.e. active presses and drug infusions) declined significantly in controls, whereas it was maintained in the mutagenized group. A progressive ratio (PR) study also showed significantly greater cumulative nicotine infusions in the PDE4B-edited group. Hence, we hypothesized that enhanced PDE4B protein activity would reduce nicotine IVSA. A positive allosteric modulator, 2-(3-(4-chloro-3-fluorophenyl)-5-ethyl-1H-1,2,4-triazol-1-yl)-N-(3,5-dichlorobenzyl)acetamide (MR-L2), was microinfused into NAcs bilaterally at FR3 or FR5; in both cohorts, MR-L2 acutely reduced nicotine IVSA. In summary, these studies show that the activity of PDE4B regulates the capacity of NAcs to maintain nicotine IVSA in face of the cost of increasing work. This finding and the results of the PR study indicate that PDE4B affects the motivation to obtain nicotine. These reverse translational studies in rats provide insight into the motivational effects of NAcs PDE4B that advance our understanding of the smoking behaviors mapped in human GWAS.

Reverse Translational Approach Using Biomaterials and Stem Cells for Intervertebral Disc Degeneration.

Reverse Translational Approach Using Biomaterials and Stem Cells for Intervertebral Disc Degeneration.

Oncogenic Activities of Tribbles1 (TRIB1) Pseudokinase Overexpressed in GBM are Mediated by Protein-Protein Interactions

Glioblastoma (GBM) is the most aggressive form of glioma with a low 5-year survival rate. The current treatments are inadequate and crippled by therapy resistance. Therefore, there is an unmet need to identify druggable therapeutic targets in GBM. In this study we identified TRIB1, a Ser/Thr pseudokinase that acts as a scaffold to initiate Ubiquitin Proteasome System-mediated degradation of its substrates. We and others have found that TRIB1 activates the canonical MAPK and Akt signaling cascades. Previous reports also suggest that TRIB1 contributes to chemotherapy resistance in various cancers. Therefore, we evaluated oncogenic roles of TRIB1 in GBM cells and its contribution to therapy resistance. Patient-centered reverse translational approach was utilized to identify novel therapeutic targets. To this end, TRIB1 was identified by statistical association (Cox regression analysis) of the patient-derived gene expression profiling data publicly available from TCGA GBM cohort. TRIB1 was functionally validated in vitro by generating stable overexpression cell lines (patient-derived) by antibiotic selection. Conditional knockdown of TRIB1 was achieved by doxycycline induction. Protein-protein interactions were evaluated by co-immunoprecipitation. Protein levels were detected by western blotting. Changes in tumor volume and overall survival (OS) were calculated. The mRNA profiling of TCGA GBM cohort revealed that increased TRIB1 gene expression was associated with worse OS of GBM patients [HR = 1.3 (1.0-1.5); P = 0.019]. The same analyses in our institutional cohort revealed a similar association. Mice bearing TRIB1 transgene overexpressing tumors had the increased tumor volume and shorter OS compared to empty vector control at the end of experiment. Overexpression of TRIB1 increased the phosphorylation/activation of ERK and Akt in patient-derived primary cell lines. Akt but not ERK activation was decreased after TRIB1 knockdown. TRIB1 bound directly to ERK and Akt in these cells. TRIB1 also formed a complex with p53, COP1 and HDAC1 in patient-derived primary cell lines. This protein-protein interaction was independent of TP53 mutation status. Our data suggest that TRIB1 overexpressed in GBM executes various oncogenic functions through interaction with different proteins. Activating ERK signaling, can induce cell proliferation. Similarly, by activating Akt it can cause prosurvival effects. Finally, by associating with HDAC1 and COP1, TRIB1 can modulate p53 function. All these protein-protein interactions ultimately contribute to chemoradiotherapy resistance in GBM cells. We are currently developing small molecule inhibitors targeting the above-mentioned interactions of TRIB1 to overcome therapeutic resistance.

Role of the tumour microenvironment in bladder cancer pathogenesis and value of the reverse translational approach: a tale of two species

AbstractBackgroundBladder cancer is a significant malignancy in humans and canines, with high death rates and devastating clinical symptoms. Despite significant advances in therapeutic management over the years, only a small fraction of bladder cancer patients respond to immunotherapies as a second line of treatment. Since immunotherapies act on various cellular and molecular targets in the bladder cancer tumour microenvironment (TME), a greater understanding of the various types of immune cells, immune checkpoint molecules and cytokines involved in antitumour immunity will help to improve the success rate of current and novel immunotherapies, therefore enhancing the patients quality of life. Accumulating evidence shows fundamental similarities between human and canine muscle‐invasive bladder cancer (MIBC) in their clinical, histological and molecular features.AimThis review focuses on comparative aspects of MIBC in both species, highlighting the role that canines can play as a model for studying MIBC. Additionally, we discuss the various types of immune cells within bladder cancer TME that can influence the prognosis and response to immunotherapy and chemotherapy in both species.ConclusionIt remains challenging to recapitulate the heterogeneity and complexity of the bladder cancer tumour microenvironment using in vivo and in vitro models, such as zebrafish and 3D organoids models. Optimization of these techniques to create a more representative translational model has been facilitated through in vitro immune‐oncology cocultures, which are considered promising tools to help select the ideal individualized treatment options and predict disease prognosis.

397-P: Epigenetic Regulation of the Chemokine CXCL2 in Acute Kidney Injury in Diabetes

People with diabetes are much more likely to develop acute kidney injury (AKI) than people who do not have diabetes. Here we took a reverse translational approach to identify factors that increase the risk of AKI in diabetes. Aged diabetic high fat diet-fed (DM-HFD) mice and nondiabetic controls were randomized to sham or bilateral renal ischemia reperfusion injury (IRI) surgery, with phenotyping performed 24h later. DM-HFD mice with IRI had increased plasma BUN and creatinine and increased tubule injury in comparison to either nondiabetic mice with IRI or DM-HFD mice that underwent sham surgery. NanoString nCounter analysis of AKI kidneys revealed that 47 of 248 inflammatory genes were differentially expressed in the kidneys of DM-HFD IRI mice in comparison to nondiabetic mice with IRI. The top hit was the chemokine Cxcl2, which was upregulated 2429-fold compared with sham-operated nondiabetic mice and 92-fold compared with nondiabetic mice with IRI. Urine CXCL2 excretion was increased >200-fold in DM-HFD IRI mice. By RNAscope in situ hybridization, Cxcl2 was observed to be upregulated in DM-HFD IRI kidneys, being localized primarily to infiltrating inflammatory cells, but also expressed in glomeruli and scattered tubule epithelial cells. Stimulation of bone marrow-derived macrophages (BMDMs) with lipopolysaccharide caused release of CXCL2 into the culture medium, with a significant increase in CXCL2 secretion by BMDMs from mice with myeloid cell deletion of the histone demethylase KDM6A (LysMCre+Kdma6fl/ymice). In summary, augmented AKI in aged diabetic high fat diet-fed mice is accompanied by upregulation of the myeloid cell chemoattractant CXCL2, which itself is under the influence of the histone demethylating enzyme KDM6A. The findings provide insights into the molecular causes of worsened AKI in diabetes and they illustrate that, like chronic kidney disease in diabetes, AKI processes in diabetes are also under epigenetic regulation. Disclosure S. Batchu: None. V. Yerra: None. Y. Hong: None. P. Plant: None. Y. Liu: None. A. Advani: None. Funding Canadian Institutes of Health Research (PJT166083 to A.A.)

A Scenario-Generic Neural Machine Translation Data Augmentation Method

Amid the rapid advancement of neural machine translation, the challenge of data sparsity has been a major obstacle. To address this issue, this study proposes a general data augmentation technique for various scenarios. It examines the predicament of parallel corpora diversity and high quality in both rich- and low-resource settings, and integrates the low-frequency word substitution method and reverse translation approach for complementary benefits. Additionally, this method improves the pseudo-parallel corpus generated by the reverse translation method by substituting low-frequency words and includes a grammar error correction module to reduce grammatical errors in low-resource scenarios. The experimental data are partitioned into rich- and low-resource scenarios at a 10:1 ratio. It verifies the necessity of grammatical error correction for pseudo-corpus in low-resource scenarios. Models and methods are chosen from the backbone network and related literature for comparative experiments. The experimental findings demonstrate that the data augmentation approach proposed in this study is suitable for both rich- and low-resource scenarios and is effective in enhancing the training corpus to improve the performance of translation tasks.

Integrating Forward and Reverse Translation in PBPK Modeling to Predict Food Effect on Oral Absorption of Weakly Basic Drugs.

Ketoconazole and posaconazole are two weakly basic broad-spectrum antifungals classified as Biopharmaceutics Classification System class II drugs, indicating that they are highly permeable, but exhibit poor solubility. As a result, oral bioavailability and clinical efficacy can be impacted by the formulation performance in the gastrointestinal system. In this work, we have leveraged in vitro biopharmaceutics and clinical data available in the literature to build physiologically based pharmacokinetic (PBPK) models for ketoconazole and posaconazole, to determine the suitability of forward in vitro-in vivo translation for characterization of in vivo drug precipitation, and to predict food effect. A stepwise modeling approach was utilized to derive key parameters related to absorption, such as drug solubility, dissolution, and precipitation kinetics from in vitro data. These parameters were then integrated into PBPK models for the simulation of ketoconazole and posaconazole plasma concentrations in the fasted and fed states. Forward in vitro-in vivo translation of intestinal precipitation kinetics for both model drugs resulted in poor predictions of PK profiles. Therefore, a reverse translation approach was applied, based on limited fitting of precipitation-related parameters to clinical data. Subsequent simulations for ketoconazole and posaconazole demonstrated that fasted and fed state PK profiles for both drugs were adequately recapitulated. The two examples presented in this paper show how middle-out modeling approaches can be used to predict the magnitude and direction of food effects provided the model is verified on fasted state PK data.

Electrophysiological signatures of reward learning in the rodent touchscreen-based Probabilistic Reward Task.

Blunted reward learning and reward-related activation within the corticostriatal-midbrain circuitry have been implicated in the pathophysiology of anhedonia and depression. Unfortunately, the search for more efficacious interventions for anhedonic behaviors has been hampered by the use of vastly different preclinical and clinical assays. In a first step in addressing this gap, in the current study, we used event-related potentials and spectral analyses in conjunction with a touchscreen version of the rodent Probabilistic Reward Task (PRT) to identify the electrophysiological signatures of reward learning in rats. We trained 11 rats (5 females and 6 males) on the rodent touchscreen-based PRT and subsequently implanted them with deep electrodes in the anterior cingulate cortex (ACC) and nucleus accumbens (NAc) for local field potentials recordings during the PRT. Behaviorally, the expected responsivity-to-reward profile was observed. At the electrophysiological level, we identified a negative amplitude deflection 250-500 ms after feedback in the ACC and NAc electrodes, as well as power increase in feedback-locked delta (1-5 Hz) and alpha/beta (9-17 Hz) bands in both electrodes for rewarded trials. Using a reverse-translational approach, we identified electrophysiological signatures of reward learning in rats similar to those described in humans. These findings and approaches might provide a useful translational platform to efficiently evaluate novel therapeutics targeting anhedonia.

Translational potential of JAX humanized APOE mice: Metabolism and body composition in very old mice

AbstractBackgroundApolipoprotein (APOE) is the strongest genetic risk factor for the development of late‐onset Alzheimer’s disease (LOAD). Clinically, LOAD exhibits strong sex effects with females being twice at the risk of developing AD than males. Models exhibiting strong AD etiology are used in preclinical settings, however lack of translatability necessitates model development with a reverse translational approach. This study explores metabolic hallmarks of LOAD assessing the clinical translatability of the JAX humanized APOE (hAPOE) mouse model.MethodhAPOE mice (M/F, n = 65, mean age = 23.35 +/‐ 0.90 months) with e3/3, e3/4 or e4/4 genotype underwent a metabolic and body composition screening assay including fasting blood glucose (FBG) and ketone (FKB) measurement, EchoMRI, and 18F‐FDG‐PET brain imaging, prior to being sacrificed. Blood plasma was analyzed for triglyceride levels. Cerebral FDG‐PET standardized uptake values were normalized to cerebellum. All data were analyzed with genotype x sex analysis of variance. P ≤ 0.05 was considered statistically significant and post‐hoc pairwise analyses were conducted for interaction effects with p ≤ 0.1 (FKB, adipose index) for future study planning.ResultFemales had significantly lower lean mass (p < 0.0001) and FBG (p=0.05), with a trend toward lower glucose uptake on FDG‐PET compared to males (p=0.072). Post‐hoc interaction analyses of FKB (interaction p=0.105) demonstrated lower ketone body levels in male e4/4s compared to female e4/4s (uncorrected p=0.0685) and male e3/4s (p=0.034). Post‐hoc interaction analyses of adipose index (interaction p=0.093) demonstrated greater adiposity in female e4/4s compared to male e4/4s (uncorrected p=0.005) and female e3/3s (uncorrected p=0.035). No effects of APOE genotype were observed.ConclusionIn a mouse cohort at a comparable human age of ∼ 70 years, lower lean mass, fasting blood glucose, and glucose uptake in the brain for female mice indicates ongoing bioenergetic deficits and is corroborated by observations within clinical populations. Greater adiposity in female e4/4s suggests that brain bioenergetic demand is not being met through fat reserves. However, the overall lack of genotypic effects suggests this bioenergetic crisis is not driven by hAPOE in this animal model, limiting clinical translatability.NIA R01AG057931; University of Arizona TBIR (NIH S10 OD025016).

TRIBBLES1 (TRIB1) Pseudokinase Induces Treatment Resistance in GBM by Promoting Survival of Tumor Cells through the Akt Pathway

<h3>Purpose/Objective(s)</h3> Glioblastoma (GBM; WHO grade IV) is the most aggressive form of glioma with a dismal 5-year survival rate. The current therapies are largely ineffective and therapy resistance is commonly encountered. Therefore, there is an urgent need to identify druggable therapeutic targets in GBM. In this study we identified TRIB1, a Ser/Thr pseudokinase that functions as a scaffold to initiate Ubiquitin Proteasome System-mediated degradation of target proteins in the cell. It has also been shown to activate the MAPK and Akt signaling cascades in various cell types. Reports also suggest that TRIB1 contributes to chemotherapy resistance in various cancers. Therefore, we evaluated the role of TRIB1 in GBM cell proliferation/ survival and its contribution to therapy resistance. <h3>Materials/Methods</h3> We utilized patient-centered reverse translational approach to identify novel therapeutic targets. To this end, TRIB1 was identified by statistical association (Cox regression analysis) of the patient-derived mRNA profiling data publicly available from TCGA GBM cohort. TRIB1 was functionally validated <i>in vitro</i> by generating stable overexpression cell lines by antibiotic selection. TRIB1 was conditionally knocked down by doxycycline induction. Co-immunoprecipitation was performed to evaluate protein-protein interactions. Protein levels were detected by western blotting. Site-directed mutagenesis was performed to mutate specific amino acid. Patient derived primary cell lines overexpressing the wild type and mutant (W337A) Trib1 transgenes were injected intracranially to create tumors in nude mice. Changes in tumor volume and overall survival (OS) were determined. <h3>Results</h3> The mRNA profiling of TCGA GBM cohort revealed that increased <i>TRIB1</i> gene expression was associated with worse OS of GBM patients (HR=1.3 (1.0-1.5); P=0.019). Mice bearing TRIB1 transgene overexpressing tumors had highest tumor volume and lowest OS whereas mice bearing TRIB1-W337A tumors had the highest OS and lower tumor volume when measured. We also observed that overexpression of TRIB1 caused a decrease in apoptosis of primary GBM cell lines after RT and TMZ treatments <i>in vitro</i>. Overexpression of TRIB1 also increased the phosphorylation/activation of Akt in patient derived primary cell lines. Akt activation was similarly decreased after TRIB1 knockdown. TRIB1 bound directly to Akt in these cells. Following PI3K inhibition, Akt phosphorylation was detected to a greater level in TRIB1 overexpressing cells. <h3>Conclusion</h3> Our data suggest that heightened expression of TRIB1 in GBM cells contributes to RT/TMZ resistance by activating the prosurvival Akt signaling cascades. Therefore, targeting TRIB1 mediated Akt activation could provide a therapeutic benefit over targeting Akt directly, which may reduce Akt's oncogenic signal without risking the toxicity associated with available Akt- specific inhibitors.

Microbially derived polyunsaturated fatty acid as a modulator of gastrointestinal motility.

Gastrointestinal (GI) motility requires coordination among several cell types in the intestinal epithelium and the neuromuscular apparatus. A disruption in GI motility was primarily attributed to disruption of this coordinated effort among different host cells, but recent studies have begun to uncover how the products of gut microbiota can alter GI motility by modulating the function of different host cells and the interactions among them. In this issue of the JCI, Chen, Qiu, et al. used a reverse translation approach, isolating a Shigella sp. — peristaltic contraction–inhibiting bacterium (PIB) — from a cohort of patients with intractable constipation. They identified an ω-3 polyunsaturated fatty acid (PUFA), docosapentaenoic acid (DPA), produced by this Shigella variant, as an important driver of constipation using a series of microbiologic, biochemical, and genetic manipulations combined with in vitro and in vivo studies. This finding advances the field, given that production of DPA is rare in the human gut and appears to have a distinct effect on GI physiology.

A reverse translational pharmacological approach to understand the underlying mechanisms of the reported association between hydrochlorothiazide and non-melanoma skin cancer.

A reverse translational pharmacological approach to understand the underlying mechanisms of the reported association between hydrochlorothiazide and non-melanoma skin cancer.

Bile acid-induced lung injury: update of reverse translational biology.

The presence of bile acids in lung tissue is associated with some clinical features observed in various medical specialties, but it took time to understand that these are due to a "bile acid-induced lung injury" since specific translational studies and cross-disciplinary awareness were lacking. We used a reverse translational approach to update and summarize the current knowledge about the mechanisms of bile acid-induced lung injury. This has been done in a cross-disciplinary fashion since these conditions may occur in patients of various ages and in different medical fields. We here define these clinical conditions, then we review the physiopathology of these conditions and the animal models used to mimic them, and, finally, their pathobiology. Mechanisms of bile acid-induced lung injury have been partially clarified over time and are represented by 1) the interaction with secretory phospholipase A2 pathway, 2) the effect on surfactant function and structure, 3) the biological effects on inflammation and local immunity, and 4) the direct cellular toxicity. These mechanisms are schematically illustrated and histological comparisons between acute respiratory distress syndrome (ARDS) induced by bile acids and other triggers are also provided. Based on these mechanisms, we propose possible direct therapeutic applications and, finally, we discuss further research steps to improve the understanding of processes that generate pathological clinical conditions.

Transforming a clinical fluorescent dye to sense and treat iron overload disorders: a new reverse translational approach in precision medicine

Transforming a clinical fluorescent dye to sense and treat iron overload disorders: a new reverse translational approach in precision medicine

Elevated Serum Purine Levels in Schizophrenia: A Reverse Translational Study to Identify Novel Inflammatory Biomarkers.

BackgroundImmunological markers and related signaling molecules in the blood are altered in schizophrenia mouse models, in acutely relapsed patients with schizophrenia, and in persons at a clinically high risk for subsequently developing psychosis, highlighting their potential as prognostic and theranostic biomarkers. Therefore, we herein aimed to identify novel potential biomarkers in the serum that are associated with purinergic signaling.MethodsTo our knowledge, this is the first study to assess the correlations among the levels of human serum adenine nucleotides (ATP, ADP), adenosine, P2X7 receptor, and disease activity in patients hospitalized due to an acute relapse of schizophrenia (n = 53) and healthy controls (n = 47). In addition, to validate these findings using a reverse translational approach, we examined the same parameters in an acute phencyclidine-induced schizophrenia mouse model.ResultsWe found consistently elevated levels of ATP, ADP, interleukin (IL)-6, and IL-10 in both schizophrenia groups compared with the controls. The levels of adenosine, IL-1β, IL-12, and C-reactive protein were also increased in the human patient samples. Moreover, ATP and ADP were significantly positively correlated with the Positive and Negative Symptom Scale item “lack of judgment and insight”; IL-1β, IL-12, and tumour necrosis factor alpha were significantly positively correlated with “tension” and “depression”; and “disorientation” and “poor attention” were correlated significantly with IL-6 and IL-8.ConclusionsOur study suggests the promising potential of blood purines and inflammatory markers as future prognostic tools.

Radiotherapy orchestrates natural killer cell dependent antitumor immune responses through CXCL8.

Radiotherapy is a mainstay cancer therapy whose antitumor effects partially depend on T cell responses. However, the role of Natural Killer (NK) cells in radiotherapy remains unclear. Here, using a reverse translational approach, we show a central role of NK cells in the radiation-induced immune response involving a CXCL8/IL-8–dependent mechanism. In a randomized controlled pancreatic cancer trial, CXCL8 increased under radiotherapy, and NK cell positively correlated with prolonged overall survival. Accordingly, NK cells preferentially infiltrated irradiated pancreatic tumors and exhibited CD56dim-like cytotoxic transcriptomic states. In experimental models, NF-κB and mTOR orchestrated radiation-induced CXCL8 secretion from tumor cells with senescence features causing directional migration of CD56dim NK cells, thus linking senescence-associated CXCL8 release to innate immune surveillance of human tumors. Moreover, combined high-dose radiotherapy and adoptive NK cell transfer improved tumor control over monotherapies in xenografted mice, suggesting NK cells combined with radiotherapy as a rational cancer treatment strategy.

Identification of Novel Diagnostic Markers for Malignant Pleural Mesothelioma Using a Reverse Translational Approach Based on a Rare Synchronous Tumor.

Although the routine use of immunohistochemistry has improved the accuracy of histopathologic diagnosis in clinical practice, new methods for discovering novel diagnostic markers are still needed. We sought new diagnostic markers for malignant pleural mesothelioma (MPM) using a reverse translational approach with limited archival tissues from a very rare case. Total RNA extracted from formalin-fixed paraffin-embedded (FFPE) tissues of a synchronous collision tumor consisting of MPM and pulmonary adenocarcinoma (PAC) was employed for gene expression profiling (GEP) analysis. Among the 54 genes selected by GEP analysis, we finally identified the following two candidate MPM marker genes: PHGDH and TRIM29. Immunohistochemical analysis of 48 MM and 20 PAC cases showed that both PHGDH and TRIM29 had sensitivity and specificity almost equivalent to those of calretinin (sensitivity 50% and 46% vs. 63%, and specificity 95% and 100% vs. 100%, respectively). Importantly, of the 23 epithelioid MMs, all 3 calretinin-negative cases were positive for TRIM29. These two markers may be diagnostically useful for immunohistochemical distinction between MPMs and PACs. This successful reverse translational approach based on FFPE samples from one very rare case encourages the further use of such samples for the development of novel diagnostic markers.