Mechanistic Support Research Articles

Temperature-dependent predation predicts a more reptilian future.

Diversity increases toward the tropics, but the strength of this pattern diverges with thermoregulatory strategy. Synthesizing over 30,000 species distributions, we quantified patterns of richness in terrestrial vertebrates, and present evidence for a latitudinal gradient of community composition. We observe a two orders of magnitude shift in comparative diversity with temperature, from endothermic mammal and avian dominance near the poles, toward ectothermic reptile and amphibian majority in the tropics. Next, we provide mechanistic support for a corresponding latitudinal gradient of predatory interactions. Using automated video tracking in >4500 trials, we show that differences in thermal sensitivity of locomotion in endothermic predators and ectothermic prey favors endotherms in colder environments and yields theoretically predicted foraging outcomes across thermal conditions, including the number of strikes, the distance traveled, and the time to capture prey. We also present evidence that endotherms use thermal cues to anticipate prey behavior, modulating the impact of temperature. Finally, we integrate theory and data to forecast future patterns of diversity, revealing that as the world get warmer, it will become increasingly reptilian. Overall, our results point toward a broad reorganization of vertebrate diversity with latitude, elevation, and temperature: from endotherm dominance in cold systems toward ectotherm dominance in warm.

Danshensu reduces neuronal excitability by enhancing potassium currents in bushy cells in the mouse cochlear nucleus.

Danshensu, also known as salvianic acid A, is a primary active compound extracted from a traditional Chinese herb Danshen (Salvia miltiorrhiza). While its antioxidative and neuroprotective effects are well-documented, the underlying mechanisms are poorly understood. In this study, we sought out to investigate if and how Danshensu modulates neuronal excitability and voltage-gated ionic currents in the central nervous system. We prepared brain slices of the mouse brainstem and performed patch-clamp recording in bushy cells in the anteroventral cochlear nucleus, with or without Danshensu incubation for 1 h. QX-314 was used internally to block Na+ current, while tetraethylammonium and 4-aminopyridine were used to isolate different subtypes of K+ current. We found that Danshensu of 100 μm decreased the input resistance of bushy cells by approximately 60% and shifted the voltage threshold of spiking positively by approximately 7 mV, resulting in significantly reduced excitability. Furthermore, we found this reduced excitability by Danshensu was caused by enhanced voltage-gated K+ currents in these neurons, including both low voltage-activated IK,A, by approximately 100%, and high voltage-activated IK,dr, by approximately 30%. Lastly, we found that the effect of Danshensu on K+ currents was dose-dependent in that no enhancement was found for Danshensu of 50 μm and Danshensu of 200 μm failed to cause significantly more enhancement on K+ currents when compared to that of 100 μm. We found that Danshensu reduced neuronal excitability in the central nervous system by enhancing voltage-gated K+ currents, providing mechanistic support for its neuroprotective effect widely seen in vivo.

TRIM33 enhances the ubiquitination of TFRC to enhance the susceptibility of liver cancer cells to ferroptosis

BackgroundHepatocellular carcinoma (HCC) is a common malignancy, and ferroptosis is a novel form of cell death driven by excessive lipid peroxidation. In recent years, ferroptosis has been widely utilized in cancer treatment, and the ubiquitination modification system has been recognized to play a crucial role in tumorigenesis and metastasis. Increasing evidence suggests that ubiquitin regulates ferroptosis-related substrates involved in this process. However, the precise mechanism of utilizing ubiquitination modification to regulate ferroptosis for HCC treatment remains unclear. MethodsIn this study, we detected the expression of TRIM33 in HCC using immunohistochemistry and western blotting techniques. The functional role of TRIM33 was verified through both in vitro and in vivo experiments. To evaluate the level of ferroptosis, mitochondrial superoxide levels, MDA levels, Fe2+ levels, and cell viability were assessed. Downstream substrates of TRIM33 were screened and confirmed via immunoprecipitation, immunofluorescence staining, and ubiquitination modification experiments. ResultsOur findings demonstrate that TRIM33 inhibits the growth and metastasis of HCC cells both in vitro and in vivo while promoting their susceptibility to ferroptosis. Mechanistically speaking, TRIM33 induces cellular ferroptosis through E3 ligase-dependent degradation of TFRC—a known inhibitor of this process—thus elucidating the specific type and site at which TFRC undergoes modification by TRIM33. ConclusionIn summary, our study reveals an important role for TRIM33 in HCC treatment while providing mechanistic support for its function. Additionally highlighted is the significance of ubiquitination modification leading to TFRC degradation—an insight that may prove valuable for future targeted therapies.

Early-life infection depletes preleukemic cells in a mouse model of hyperdiploid B-cell acute lymphoblastic leukemia

AbstractEpidemiological studies report opposing influences of infection on childhood B-cell acute lymphoblastic leukemia (B-ALL). Although infections in the first year of life appear to exert the largest impact on leukemia risk, the effect of early pathogen exposure on the fetal preleukemia cells (PLC) that lead to B-ALL has yet to be reported. Using cytomegalovirus (CMV) infection as a model early-life infection, we show that virus exposure within 1 week of birth induces profound depletion of transplanted E2A-PBX1 and hyperdiploid B-ALL cells in wild-type recipients and in situ–generated PLC in Eμ-ret mice. The age-dependent depletion of PLC results from an elevated STAT4-mediated cytokine response in neonates, with high levels of interleukin (IL)-12p40–driven interferon (IFN)-γ production inducing PLC death. Similar PLC depletion can be achieved in adult mice by impairing viral clearance. These findings provide mechanistic support for potential inhibitory effects of early-life infection on B-ALL progression and could inform novel therapeutic or preventive strategies.

Heart rate variability analysis in obstructive sleep apnea patients with daytime sleepiness.

Recent studies suggest that sleepy patients with obstructive sleep apnea (OSA) are at higher risk for incident cardiovascular disease. This study assessed cardiac autonomic function in sleepy versus non-sleepy patients with OSA using heart rate variability (HRV) analysis. We hypothesized that HRV profiles of sleepy patients would indicate higher cardiovascular risk. Electrocardiograms (ECG) derived from polysomnograms (PSG) collected by the Sydney Sleep Biobank were used to study HRV in groups of sleepy (ESS ≥ 10) and non-sleepy OSA patients (ESS < 10). HRV parameters were averaged across available ECG signals during N2 sleep. A total of 421 patients were evaluated, with a mean age of 54 (14) years, body mass index of 33 (9) kg/m2, apnea-hypopnea index of 21 (28) events/h, and 66% male. The sleepy group consisted of 119 patients and the non-sleepy group 302 patients. Sleepy patients exhibited lower HRV values for: root mean square successive difference (RMSSD, p = 0.028), total power (TP, p = 0.031), absolute low frequency (LF, p = 0.045), and high-frequency (HF, p = 0.010) power compared to non-sleepy patients. Sleepy patients with moderate-to-severe OSA exhibited lower HRV values for: (RMSSD, p = 0.045; TP, p = 0.052), absolute LF (p = 0.051), and HF power (p = 0.025). There were no differences in other time and frequency domain HRV markers. This study shows a trend toward parasympathetic withdrawal in sleepy OSA patients, particularly in moderate-to-severe cases, lending mechanistic support to the link between the sleepy phenotype and CVD risk in OSA.

Abstract 1786: Metabolic rewiring induced by protein or calorie restriction provides unique therapeutic vulnerabilities in translocation and select clear cell renal cell carcinomas

Abstract While the approval of several combinations with immune checkpoint inhibitors have significantly improved the outcome of kidney cancer patients, advanced clear cell and non-clear cell renal cell carcinoma (RCC) remain a clinical challenge. There has been a longstanding interest in using dysregulated metabolism as a therapeutic target due to its association with the pathogenesis and progression of several cancer types. As one of these highly metabolic tumors, RCC has the potential to greatly benefit from strategies targeting tumor metabolism. Dietary interventions are an innovative adjunct therapy that has the potential to induce a wide range of anti-cancer effects by disrupting metabolism due to the role diet plays in providing tumors with macromolecules for energy metabolism. Unfortunately, tumor cell plasticity, intratumor heterogeneity, and a lack of understanding regarding metabolic vulnerabilities in tumors limit the efficacy of implementing these strategies and a fundamental gap in the field remains as to how basic metabolic requirements of the tumor can be targeted with specific dietary interventions. In this study, the inherent metabolic phenotypes of translocation RCC (tRCC) and clear cell RCC (ccRCC) were characterized by integrating in vivo metabolomics and transcriptomics data, assessing mitochondrial and glycolytic activity in vitro via Seahorse, and performing immunohistochemical analyses of patient samples. Interestingly, the respective mitochondrial and glycolytic dependencies in tRCC and ccRCC patient-derived xenograft (PDX) models appear to influence the response to nutrient deprivation as seen by in vitro assays conducted after amino acid or glucose restriction. Importantly, these results indicate that dietary interventions may be able to selectively target metabolic vulnerabilities in tRCC and ccRCC. Ongoing combination studies with either protein or calorie restriction and targeted therapies, such as standard of care receptor tyrosine-kinase inhibitors, suggest the benefit of using dietary interventions as an adjunct therapy in RCC. Overall, these studies reveal that selective dietary interventions alter the metabolic landscape of RCC and expose vulnerabilities that can be exploited with targeted therapy. As dietary interventions are a safe, affordable, and potentially well tolerated adjunct therapy, we anticipate that the results of these studies will provide the biologic and mechanistic support for translating this strategy to advanced RCC patients. Citation Format: Sabrina A. Orsi, Christopher Rupert, Sean Colligan, Musea Chang, Abbas Jawadwala, Ilaria Delle Fontane, Filomena de Nigris, Roberto Pili. Metabolic rewiring induced by protein or calorie restriction provides unique therapeutic vulnerabilities in translocation and select clear cell renal cell carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1786.

Assessment of the genotoxicity of tert-butyl alcohol in an in vivo thyroid comet assay.

Chronic exposure to high (20,000 ppm) concentrations of tert-butyl alcohol (TBA) in drinking water, equivalent to ~2100 mg/kg bodyweight per day, is associated with slight increases in the incidence of thyroid follicular cell adenomas and carcinomas in mice, with no other indications of carcinogenicity. In a recent toxicological review of TBA, the U.S. EPA determined that the genotoxic potential of TBA was inconclusive, largely based on non-standard studies such as in vitro comet assays. As such, the potential role of genotoxicity in the mode of action of thyroid tumors and therefore human relevance was considered uncertain. To address the potential role of genotoxicity in TBA-associated thyroid tumor formation, CD-1 mice were exposed up to a maximum tolerated dose of 1500 mg/kg-day via oral gavage for two consecutive days and DNA damage was assessed with the comet assay in the thyroid. Blood TBA levels were analyzed by headspace GC-MS to confirm systemic tissue exposure. At study termination, no significant increases (DNA breakage) or decreases (DNA crosslinks) in %DNA tail were observed in TBA exposed mice. In contrast, oral gavage of the positive control ethyl methanesulfonate significantly increased %DNA tail in the thyroid. These findings are consistent with most genotoxicity studies on TBA and provide mechanistic support for non-linear, threshold toxicity criteria for TBA. While the mode of action for the thyroid tumors remains unclear, linear low dose extrapolation methods for TBA appear more a matter of policy than science.

An assay for aryl radicals using BHAS coupling.

Aryl radicals are intermediates in many reactions, but determining their presence unambiguously is often challenging. As we recently reported, reaction of 2-iodo-1,3-dimethylbenzene (7) in benzene with KOtBu and a suitable organic additive, leads to a base-induced homolytic aromatic substitution (BHAS) coupling reaction giving 2,6-dimethylbiphenyl (12) and biphenyl (3) as coupled products, together with xylene (13). In this case, biphenyl arises from a radical translocation and is the major coupling product. This paper now quantitatively investigates that reaction, which shows a very similar ratio for 3 : 12 [ca. 4 : 1] when using different sources of radical initiation. Deuterium isotope studies provide detailed mechanistic support for the proposed mechanism; when carried out in C6D6vs. C6H6, the reaction is characterised by a strong isotope effect for formation of 3-d10vs. 3, but not for formation of 12-d5vs. 12. These distinctive properties mean that the transformation can act as an assay for aryl radicals. An advantage of such a BHAS process is its sensitivity, since it involves a chain reaction that can amplify radical activity.

Response of stomatal conductance, transpiration, and photosynthesis to light and CO2 for rice leaves with different appearance days.

To investigate the dynamics of stomata, transpiration, and photosynthesis under varying light intensities and CO2 conditions during leaf development, the light response and CO2 response of stomatal conductance (g sw), transpiration rate (T r), and net photosynthetic rate (P n) were observed for rice leaves at different days after leaf emergence (DAE). The results showed that (1) as photosynthetically active radiation (PAR) increased, leaf g sw, T r, and P n initially increased rapidly and linearly, followed by a more gradual rise to maximum values, and then either stabilized or showed a declining trend. The maximum g sw, T r, and P n were smaller and occurred earlier for old leaves than for young leaves. The g sw, T r, and P n all exhibited a linear decreasing trend with increasing DAE, and the rate of decrease slowed down with the reduction in PAR; (2) as the CO2 concentration (C a) increased, g sw and T r decreased gradually to a stable minimum value, while P n increased linearly and slowly up to the maximum and then kept stable or decreased. The g sw, T r, and P n values initially kept high and then decreased with the increase of DAE. These results contribute to understanding the dynamics in g sw, T r, and P n during rice leaf growth and their response to varied light and CO2 concentration conditions and provide mechanistic support to estimate dynamic evapotranspiration and net ecosystem productivity at field-scale and a larger scale in paddy field ecosystems through the upscaling of leaf-level stomatal conductance, transpiration, and photosynthesis.

Modifying Alzheimer's disease pathophysiology with photobiomodulation: model, evidence, and future with EEG-guided intervention.

This manuscript outlines a model of Alzheimer's Disease (AD) pathophysiology in progressive layers, from its genesis to the development of biomarkers and then to symptom expression. Genetic predispositions are the major factor that leads to mitochondrial dysfunction and subsequent amyloid and tau protein accumulation, which have been identified as hallmarks of AD. Extending beyond these accumulations, we explore a broader spectrum of pathophysiological aspects, including the blood-brain barrier, blood flow, vascular health, gut-brain microbiodata, glymphatic flow, metabolic syndrome, energy deficit, oxidative stress, calcium overload, inflammation, neuronal and synaptic loss, brain matter atrophy, and reduced growth factors. Photobiomodulation (PBM), which delivers near-infrared light to selected brain regions using portable devices, is introduced as a therapeutic approach. PBM has the potential to address each of these pathophysiological aspects, with data provided by various studies. They provide mechanistic support for largely small published clinical studies that demonstrate improvements in memory and cognition. They inform of PBM's potential to treat AD pending validation by large randomized controlled studies. The presentation of brain network and waveform changes on electroencephalography (EEG) provide the opportunity to use these data as a guide for the application of various PBM parameters to improve outcomes. These parameters include wavelength, power density, treatment duration, LED positioning, and pulse frequency. Pulsing at specific frequencies has been found to influence the expression of waveforms and modifications of brain networks. The expression stems from the modulation of cellular and protein structures as revealed in recent studies. These findings provide an EEG-based guide for the use of artificial intelligence to personalize AD treatment through EEG data feedback.

Efficient and eco-friendly cadmium ion recycling: Ultrasonic enhancement of aluminum powder replacement for low-temperature industrial applications

Replacing cadmium ions in cadmium-containing solutions with aluminum powder is beneficial for cadmium resource recycling and environmental protection. However, the conventional aluminum powder replacement method requires harsh temperatures and prolonged conditions. In this study, the effect and mechanism of ultrasound on the replacement of cadmium with aluminum powder were investigated at low temperatures. Ultrasound has been proven to promote the etching of alumina films through the use of TEM and XPS, providing mechanistic support for the superiority of the new process. A degree of Cd replacement as high as 95.08 % is achieved at a low temperature (60 ℃) and in a short time (20 min) when using ultrasonicated aluminum powder replacement, which is 42.17 % higher than that of conventional aluminum powder. Compared with conventional aluminum powder replacement conditions with the same effect, the introduction of ultrasound can reduce the temperature by 30℃ and shorten the replacement time by 2/3, which has significant advantages in reaction efficiency and safety. The strengthening mechanism of ultrasound on the replacement effect of aluminum powder at low temperatures is revealed through detailed discussions on the corrosion of alumina films, agglomeration of aluminum powder, and adhesion of replacement products to the surface of aluminum powder, dissolved oxygen in the solution, and redissolution of cadmium. Therefore, a new approach for replacing aluminum powder in solutions with high Cd2+ concentrations at low temperatures is proposed in this work, which is expected to solve the existing harsh and dangerous problems of industrial aluminum powder replacement.

Obesity worsens mitochondrial quality control and does not protect against skeletal muscle wasting in murine cancer cachexia.

More than 650 million people are obese (BMI>30) worldwide, which increases their risk for several metabolic diseases and cancer. While cachexia and obesity are at opposite ends of the weight spectrum, leading many to suggest a protective effect of obesity against cachexia, mechanistic support for obesity's benefit is lacking. Given that obesity and cachexia are both accompanied by metabolic dysregulation, we sought to investigate the impact of obesity on skeletal muscle mass loss and mitochondrial dysfunction in murine cancer cachexia. Male C57BL/6 mice were given a purified high fat or standard diet for 16weeks before being implanted with 106 Lewis lung carcinoma (LLC) cells. Mice were monitored for 25days, and hindlimb muscles were collected for cachexia indices and mitochondrial assessment via western blotting, high-resolution respirometry and transmission electron microscopy (TEM). Obese LLC mice experienced significant tumour-free body weight loss similar to lean (-12.8% vs. -11.8%, P=0.0001) but had reduced survival (33.3% vs. 6.67%, χ2 =10.04, P=0.0182). Obese LLC mice had reduced muscle weights (-24%, P<0.0354) and mCSA (-16%, P=0.0004) with similar activation of muscle p65 (P=0.0337), and p38 (P=0.0008). ADP-dependent coupled respiration was reduced in both Obese and Obese LLC muscle (-30%, P=0.0072) consistent with reductions in volitional cage activity (-39%, P<0.0001) and grip strength (-41%, P<0.0001). TEM revealed stepwise reductions in intermyofibrillar and subsarcolemmal mitochondrial size with Obese (IMF: -37%, P=0.0009; SS: -21%, P=0.0101) and LLC (IMF: -40%, P=0.0019; SS: -27%, P=0.0383) mice. Obese LLC mice had increased pAMPK (T172; P=0.0103) and reduced FIS1 (P=0.0029) and DRP1 (P<0.0001) mitochondrial fission proteins, which were each unchanged in Lean LLC. Further, mitochondrial TEM analysis revealed that Obese LLC mice had an accumulation of damaged and dysfunctional mitochondria (IMF: 357%, P=0.0395; SS: 138%, P=0.0174) in concert with an accumulation of p62 (P=0.0328) suggesting impaired autophagy and clearance of damaged mitochondria. Moreover, we observed increases in electron lucent vacuoles only in Obese LLC muscle (IMF: 421%, P=0.0260; SS: 392%, P=0.0192), further supporting an accumulation of damaged materials that cannot be properly cleared in the obese cachectic muscle. Taken together, these results demonstrate that obesity is not protective against cachexia and suggest exacerbated impairments to mitochondrial function and quality control with a particular disruption in the removal of damaged mitochondria. Our findings highlight the need for consideration of the severity of obesity and pre-existing metabolic conditions when determining the impact of weight status on cancer-induced cachexia and functional mitochondrial deficits.

Circadian epidemiology: Structuring circadian causes of disease and practical implications

ABSTRACT That disruptions of the body’s internal clockwork can lead to negative health consequences, including cancer, is a plausible hypothesis. Yet, despite strong mechanistic and animal support, the International Agency for Research on Cancer (IARC) experts considered epidemiological evidence as limited regarding the carcinogenicity of “shift-work involving circadian disruption” (2007) and “night shift work” (2019). We use directed acyclic graphs (DAGs) to outline a concept of circadian causes that discloses challenges when choosing appropriate exposure variables. On this basis, we propose to move beyond shift-work alone as a direct cause of disease. Instead, quantifying chronodisruption as individual doses can lead to interpretable circadian epidemiology. The hypothesis is that doses of chronodisruption cause disrupted circadian organisation by leading to desynchronization of circadian rhythms. Chronodisruption can be conceptualized as the split physiological nexus of internal and external times. Biological (or internal) night – an individual’s intrinsically favoured sleep time window – could be the backbone of circadian epidemiology. In practice, individual doses that cause disrupted circadian organisation are derived from the intersection of time intervals of being awake and an individual’s biological night. After numerous studies counted work shifts, chronobiology may now advance circadian epidemiology with more specific dose estimation – albeit with greater challenges in measurement (time-dependent individual data) and analysis (time-dependent confounding).

Alternative Pathway Masp-3 Inhibitor OMS906 Effectively and Potently Inhibits Complement-Mediated Hemolysis in Preclinical Models Mechanistically Similar to Paroxysmal Nocturnal Hemoglobinuria

Introduction: Mannan-binding lectin-associated serine protease-3 (MASP-3) activates pro-complement Factor D (CFD) to form mature CFD, a rate-limiting enzyme in the alternative pathway (AP) of the complement system. CFD controls cleavage of complement Factor B (CFB) to form AP C3 convertase, which drives an amplification loop of protease complexes that activates both C3b-mediated opsonization and the terminal pathway. OMS906 is a humanized IgG4 monoclonal antibody (mAb) that binds to and inhibits MASP-3, thereby blocking maturation of CFD and downstream AP activity. Data from mice and monkeys demonstrated that OMS906 is highly selective and effectively inhibits AP activity. MASP-3 inhibition could provide therapeutic benefit in a variety of AP-mediated diseases, such as paroxysmal nocturnal hemoglobinuria (PNH). In PNH, a rare and life-threatening disorder, red blood cells (RBCs) lacking CD59 and CD55 regulatory surface proteins are targeted for rapid clearance via intravascular hemolysis and extravascular hemolysis, due to AP dysregulation. Clinical efficacy of OMS906 in a study of patients with PNH has been reported. To provide mechanistic support, we conducted in vitro and in vivo studies using preclinical models of complement-mediated hemolysis that have mechanistic overlap with PNH. Methods: In vitro potency of OMS906 was assessed using RBCs deficient in CD55 and CD59 to mimic the physiological effect of PNH. Healthy human donor RBCs were treated with inhibitory CD55 and CD59 antibodies, then incubated under AP assay conditions with human serum (diluted to 50%) containing OMS906, anti-C5 IgG4 mAb, or an isotype mAb control. The potency of OMS906, expressed as the half-maximal inhibitory concentration (IC 50), was assessed based on prevention of hemolysis of the PNH-like RBCs in vitro. Lysis was quantified by measuring hemoglobin released into sample supernatants using spectrophotometric absorbance. The in vivo effect of OMS906 on survival of murine RBCs prone to rapid clearance was assessed using RBCs from Crry -/- mice, which lack a rodent-specific complement regulatory protein that blocks AP activation. C57BL/6J male mice were intravenously injected with fluorescent labelled Crry -/- RBCs and received OMS906 or isotype mAb control via subcutaneous injection, or anti-CFB mAb via intraperitoneal injection. Blood samples were taken daily until Day 14 and the remaining number of Crry -/- RBCs measured by flow cytometry. Results: In vitro, OMS906 inhibited terminal cell lysis in human PNH-like RBCs more potently than a C5 inhibitor. Comparison of representative drug concentration-response showed that the average functional potency was 58-fold greater with OMS906 (IC 50: 0.8 μg/mL; 3 experiments; 12 total RBC donors) than with anti-C5 mAb (IC 50: 46.2 μg/mL; 2 experiments; 4 total RBC donors). In addition, the maximal extent to which lysis was inhibited was consistently greater with OMS906 than with anti-C5 mAb. In vivo, the rate of decline in number of Crry -/- RBCs was significantly slowed in mice treated with OMS906, in contrast to the rapid loss observed in isotype mAb control-treated mice ( Figure). OMS906 recipients showed a rate of Crry -/- RBC survival similar to the anti-CFB mAb group. Conclusion: OMS906 inhibited RBC lysis more potently than a C5 inhibitor, providing evidence that MASP-3 inhibition blocks downstream terminal activity and thus intravascular hemolysis. In addition, OMS906 improved survival of Crry -/- RBCs comparably to a CFB inhibitor, indicating that MASP-3 inhibition prevents the AP-mediated destruction of RBCs predictive of extravascular hemolysis. OMS906 is currently in clinical development for the treatment of PNH.

The Combination of Lanraplenib, a Selective SYK Inhibitor, and Gilteritinib, a FLT3 Inhibitor, Targets Aberrant Proliferation and Differentiation Blockade in Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) is an aggressive disease characterized by uncontrolled clonal proliferation of poorly differentiated myeloid progenitor cells in the bone marrow and blood. Deregulated spleen tyrosine kinase (SYK) impairs myeloid differentiation and has been implicated in the pathogenesis of AML driven by FMS-like tyrosine kinase 3 (FLT3) internal tandem duplication (ITD) mutation (FLT3-ITD). While SYK is rarely mutated, it is highly activated in FLT3-ITD-mutated AML and directly phosphorylates FLT3-ITD, resulting in aberrant expression of multiple oncogenic pathways. Lanraplenib (LANRA) is a selective, next-generation, SYK inhibitor that has shown favorable PK, PD, and safety in healthy volunteers and patients with autoimmune diseases. LANRA is currently being evaluated in combination with gilteritinib, a FLT3 inhibitor, in patients with relapsed or refractory (R/R) FLT3-mutated AML (NCT05028751). By jointly inhibiting the leukemogenic network created by SYK and mutant FLT3, LANRA and gilteritinib potentially provide a new treatment regimen in a patient population with few treatment options. Method: AML cell viability in response to LANRA, gilteritinib or their combination was assessed by CellTiter Glo (CTG) at 5 days. Apoptosis was evaluated by measuring Annexin V and propidium iodide staining by flow cytometry. Myeloid differentiation was assessed by flow cytometry using CD11b and CD14 cell surface markers and/or immunohistochemistry. In addition, RNA-seq was performed to evaluate differential changes in gene expression in response to LANRA. Gene set enrichment analysis (GSEA) was performed to evaluate perturbation in leukemogenic signaling pathways. In vivo studies were conducted using the MV4;11 cell line-derived xenograft model. LANRA was administered twice daily at a dose of 75 mg/kg via subcutaneous injections, either alone or in combination with gilteritinib at a dose of 2 mg/kg dosed once daily by oral gavage, for 28 days. Results: Treatment of FLT3-ITD mutated AML cells with LANRA results in dose- and time-dependent increase in myeloid differentiation and reductions in leukemic cell viability. RNA-seq analysis of MV4;11 cells revealed that LANRA abrogates multiple downstream FLT3-ITD leukemogenic signaling pathways including JAK/STAT3/5, MAPK, mTOR and OXPHOS. Combined pharmacologic inhibition of SYK with LANRA and FLT3-ITD with gilteritinib results in robust antileukemic effects in preclinical models of FLT3 ITD-driven AML. Additionally, the combination of LANRA and gilteritinib demonstrated increased differentiation and apoptosis induction in vitro. LANRA in combination with gilteritinib showed significant tumor growth inhibition compared to either single agent in a subcutaneous FLT3-ITD AML cell line-derived xenograft mouse model. In line with these observations, a retrospective analysis of bone marrow-engrafted AML cells from a disseminated FLT3-ITD AML patient-derived xenograft mouse model showed evidence of increased myeloid differentiation and inhibition of cell proliferation. Conclusion: In preclinical models, LANRA demonstrates compelling anti-leukemic activity in combination with gilteritinib. By jointly targeting hyperproliferation and differentiation blockade, LANRA and gilteritinib may provide a new therapeutic approach that can be exploited in AML. Additionally, LANRA disrupts downstream signaling of key pathways often associated with resistance to gilteritinib, providing additional mechanistic support for continued testing of LANRA in combination with gilteritinib in FLT3-ITD-mutated AML patients. These studies helped motivate the ongoing clinical combination with gilteritinib in patients with (R/R) FLT3-mutated AML (NCT05028751).

Synthetic Chemicals: What We Have Learned and Still Need to Learn About Their Associations with Childhood Allergy and Asthma.

Prenatal and childhood exposure to synthetic chemicals, such as phenols and phthalates, have been linked to asthma and allergy, but the extent of this association and the underlying mechanisms are not fully understood. Here we provide an up-to-date review of the evidence linking phenol and phthalate exposure with childhood asthma and allergy and of proposed mechanistic pathways. Five experimental and 12 epidemiological studies that examined associations between exposures to synthetic chemicals to asthma and allergic diseases were included. An additional 14 studies provided mechanistic support for the importance of immune modification through epigenetic regulation, induction of pro-allergic T2 expression, and endocrine disruption. While recent studies have provided further experimental and epidemiological evidence for how these chemical exposures may induce childhood asthma and allergy, the recent literature remains limited. However, emerging mechanistic studies have identified chemical-induced alterations in DNA methylation of genes implicated in allergic inflammation and endocrine disruption as potential pathways. In addition, barriers to decrease exposure to synthetic chemicals at the individual level (facilitated through education) and areas for further action at the organizational and governmental levels are suggested. The latter includes transferring some of the onus from the individual to organizations and legislation to restrict marketing and access to products containing potentially harmful chemicals and provide alternative products. We also suggest future research that focuses on further elucidating pathways between exposure to disease development and identifying strategies to reduce exposure at the population level.

Divergent amplifications of CYP9A cytochrome P450 genes provide two noctuid pests with differential protection against xenobiotics

Here, we provide mechanistic support for the involvement of the CYP9A subfamily of cytochrome P450 monooxygenases in the detoxification of host plant defense compounds and chemical insecticides in Spodoptera exigua and Spodoptera frugiperda. Our comparative genomics shows that a large cluster of CYP9A genes occurs in the two species but with significant differences in its contents, including several species-specific duplicates and substantial sequence divergence, both between orthologs and between duplicates. Bioassays of CRISPR-Cas9 knockouts of the clusters show that, collectively, the CYP9As can detoxify two furanocoumarin plant defense compounds (imperatorin and xanthotoxin) and insecticides representing three different chemotypes (pyrethroids, avermectins, and oxadiazines). However, in vitro metabolic assays of heterologously expressed products of individual genes show several differences between the species in the particular CYP9As with activities against these compounds. We also find that the clusters show tight genetic linkage with high levels of pyrethroid resistance in field strains of the two species. We propose that their divergent amplifications of the CYP9A subfamily have not only contributed to the development of the broad host ranges of these species over long evolutionary timeframes but also supplied them with diverse genetic options for evolving resistance to chemical insecticides in the very recent past.

Relationship between net primary productivity and stand age in typical plantation forests in China.

Plantation forests play an important role in carbon sink in terrestrial ecosystems. Based on tree rings of five main plantation tree species (Robinia pseudoacacia, Quercus variabilis, Cunninghamia lanceolata, Pinus sylvestris var. mongolica, and Pinus tabuliformis) at 25 sites in China, we calculated the average annual NPP of standard trees in each study area by the biomass equations and extended to the stand scale. The relationships between NPP and stand age were fitted by the InTEC and Law models. The results showed that NPP of R. pseu-doacacia, C. lanceolata, and P. tabuliformis plantations increased to a peak and then leveling off with stand age, while that of Q. variabilis and P. sylvestris var. mongolica plantations reached a peak and then showed a decreasing trend. The inflection points of NPP-stand age curve for different planatations was 11 years for P. sylvestris var. mongolica, 14 years for C. lanceolata, 16 years for P. tabuliformis, and 20 years for R. pseudoacacia. The NPP peak was 6.65, 7.58, 4.70 and 2.59 t·hm-2·a-1, respectively. Both the InTEC and Law NPP-stand age models had high fitting accuracy at a large scale, with the lowest R2 of 0.95 and RMSE of 0.55 t·hm-2·a-1 for the P. sylvestris var. mongolica InTEC model and the highest R2 of 0.99 and RMSE of 0.26 t·hm-2·a-1 for the C. lanceolata InTEC model. The construction of NPP-stand age relationship for major plantations in China provided mechanistic support for the estimation of carbon sinks in plantations at long time scales and provided a reference for the diversification of afforestation tree species selection.

Warm oceans exacerbate Chinook salmon bycatch in the Pacific hake fishery driven by thermal and diel depth‐use behaviours

AbstractFisheries bycatch impacts marine species globally and understanding the underlying ecological and behavioural mechanisms could improve bycatch mitigation and forecasts in novel conditions. Oceans are rapidly warming causing shifts in marine species distributions with unknown, but likely, bycatch consequences. We examined whether thermal and diel depth‐use behaviours influenced bycatch of a keystone species (Chinook salmon; Oncorhynchus tshawytscha, Salmonidae) in the largest fishery on the US West Coast (Pacific hake; Merluccius productus, Merlucciidae) with annual consequences in a warming ocean. We used Generalized Additive Models with 20 years of data including 54,509 hauls from the at‐sea hake fishery spanning Oregon and Washington coasts including genetic information for five salmon populations. Our results demonstrate that Chinook salmon bycatch rates increased in warm ocean years explained by salmon depth‐use behaviours. Chinook salmon typically occupy shallower water column depths compared to hake. However, salmon moved deeper when sea surface temperatures (SSTs) were warm and at night, which increased overlap with hake and exacerbated bycatch rates. We show that night fishing reductions (a voluntary bycatch mitigation strategy) are effective in reducing salmon bycatch in cool SSTs by limiting fishing effort when diel vertical movements bring salmon deeper but becomes less effective in warm SSTs as salmon seek deeper thermal refugia during the day. Thermal and diel behaviours were more pronounced in southern compared with northern salmon populations. We provide mechanistic support that climate change may intensify Chinook salmon bycatch in the hake fishery and demonstrate how an inferential approach can inform bycatch management in a changing world.

Chronic alcohol exposure alters action control via hyperactive premotor corticostriatal activity

Alcohol use disorder (AUD) alters decision-making control over actions, but disruptions to the responsible neural circuit mechanisms are unclear. Premotor corticostriatal circuits are implicated in balancing goal-directed and habitual control over actions and show disruption in disorders with compulsive, inflexible behaviors, including AUD. However, whether there is a causal link between disrupted premotor activity and altered action control is unknown. Here, we find that mice chronically exposed to alcohol (chronic intermittent ethanol [CIE]) showed impaired ability to use recent action information to guide subsequent actions. Prior CIE exposure resulted in aberrant increases in the calcium activity of premotor cortex (M2) neurons that project to the dorsal medial striatum (M2-DMS) during action control. Chemogenetic reduction of this CIE-induced hyperactivity in M2-DMS neurons rescued goal-directed action control. This suggests a direct, causal relationship between chronic alcohol disruption to premotor circuits and decision-making strategy and provides mechanistic support for targeting activity of human premotor regions as a potential treatment in AUD.