Whole-body Model Research Articles

Tailoring Solution Accuracy for Fast Whole-Body Model Predictive Control of Legged Robots

Tailoring Solution Accuracy for Fast Whole-Body Model Predictive Control of Legged Robots

Abstract A018: Endothelial pyrimidine synthesis deficiency promotes tumor growth

Abstract The tumor microenvironment plays a crucial role in cancer progression and treatment response. Systemically administered metabolic cancer therapies target not only malignant but also stromal cells, including immune cells - the only stromal population previously addressed in this context. However, how such treatments impact other non-malignant cell types in tumors remains poorly understood. Here we show that inhibition of de novo pyrimidine synthesis in endothelial cells accelerates tumor growth and alters tumor immune repertoire. We found that whole-body de novo pyrimidine synthesis deficiency in mice, caused by the inducible whole-body ablation of DHODH, accelerates the growth rate of orthotopic lung tumors. Single-cell transcriptomic analysis of tumor-bearing lungs revealed that DHODH deficiency in the stroma impacts multiple cell populations, including immune and, surprisingly, endothelial cells. To explore the endothelial-specific effects, we generated a mouse model with inducible DHODH deficiency restricted to endothelial cells. The endothelium-specific model recapitulates the accelerated lung tumor growth observed in the whole-body DHODH deficiency model. Single-cell transcriptomics analysis of tumor-bearing lungs in the endothelium-specific model pointed to changes in the immune repertoire, particularly an enrichment of monocytes. We confirmed these results on the protein level using spectral flow cytometry, and we are currently in the process of uncovering the mechanism by which endothelial deficiency of pyrimidine synthesis affects the immune landscape of tumors. Inhibitors of pyrimidine de novo synthesis have been tested in clinical trials but failed due to their low efficiency. Our findings indicate that systemic treatment targeting pyrimidine synthesis may be hampered by its pro-tumorigenic effects in the endothelium, highlighting the unexpected role of endothelial metabolism in this context. Citation Format: Petra Hyrossova, Isidora Milisav, Silvia Novais, Mirko Milosevic, Jakub Rohlena, Katerina Rohlenova. Endothelial pyrimidine synthesis deficiency promotes tumor growth [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr A018.

Local depletion of large molecule drugs due to target binding in tissue interstitial space.

Drug-target binding determines a drug's pharmacodynamics but can also have a profound impact on a drug's pharmacokinetics, known as target-mediated drug disposition (TMDD). TMDD models describe the influence of drug-target binding and target turnover on unbound drug concentrations and are frequently used for biologics and drugs with nonlinear plasma pharmacokinetics. For drug targets expressed in tissues, the effect of TMDD may not be detected when analyzing plasma concentration curves, but it might still affect tissue concentrations and occupancy. This review aimed to investigate the likeliness of such a scenario by reviewing the literature for a typical range of TMDD parameter values and their impact on local drug concentrations and target occupancy in a whole-body PBPK model with TMDD. Our analysis demonstrated that tissue drug concentrations are impacted and significantly depleted in many physiological scenarios. In contrast, the effect on plasma concentrations is much lower, specifically for smaller organs with lower perfusion. Moreover, in scenarios with fast internalization of the drug-target complex, the distribution of large molecules from plasma to tissue interstitial space emerges as a rate-limiting step for the drug-target interaction. These factors may lead to overpredicting local drug concentrations when considering only plasma pharmacokinetics. A sensitivity analysis revealed the high and not always intuitive impact of drug-specific parameters, including the drug molecule hydrodynamic radius, dissociation constant (Kd), drug-target complex internalization rate constant (kint), and target dissociation rate constant (koff), on the drug's pharmacokinetics. Our analysis demonstrated that tissue TMDD needs to be considered even if plasma pharmacokinetics are linear.

Molecular basis of neurodegeneration in a mouse model of Polr3-related disease

Pathogenic variants in subunits of RNA polymerase (Pol) III cause a spectrum of Polr3-related neurodegenerative diseases including 4H leukodystrophy. Disease onset occurs from infancy to early adulthood and is associated with a variable range and severity of neurological and non-neurological features. The molecular basis of Polr3-related disease pathogenesis is unknown. We developed a postnatal whole-body mouse model expressing pathogenic Polr3a mutations to examine the molecular mechanisms by which reduced Pol III transcription results primarily in central nervous system phenotypes. Polr3a mutant mice exhibit behavioral deficits, cerebral pathology and exocrine pancreatic atrophy. Transcriptome and immunohistochemistry analyses of cerebra during disease progression show a reduction in most Pol III transcripts, induction of innate immune and integrated stress responses and cell-type-specific gene expression changes reflecting neuron and oligodendrocyte loss and microglial activation. Earlier in the disease when integrated stress and innate immune responses are minimally induced, mature tRNA sequencing revealed a global reduction in tRNA levels and an altered tRNA profile but no changes in other Pol III transcripts. Thus, changes in the size and/or composition of the tRNA pool have a causal role in disease initiation. Our findings reveal different tissue- and brain region-specific sensitivities to a defect in Pol III transcription.

Molecular basis of neurodegeneration in a mouse model of Polr3-related disease.

Pathogenic variants in subunits of RNA polymerase (Pol) III cause a spectrum of Polr3-related neurodegenerative diseases including 4H leukodystrophy. Disease onset occurs from infancy to early adulthood and is associated with a variable range and severity of neurological and non-neurological features. The molecular basis of Polr3-related disease pathogenesis is unknown. We developed a postnatal whole-body mouse model expressing pathogenic Polr3a mutations to examine the molecular mechanisms by which reduced Pol III transcription results primarily in central nervous system phenotypes. Polr3a mutant mice exhibit behavioral deficits, cerebral pathology and exocrine pancreatic atrophy. Transcriptome and immunohistochemistry analyses of cerebra during disease progression show a reduction in most Pol III transcripts, induction of innate immune and integrated stress responses and cell-type-specific gene expression changes reflecting neuron and oligodendrocyte loss and microglial activation. Earlier in the disease when integrated stress and innate immune responses are minimally induced, mature tRNA sequencing revealed a global reduction in tRNA levels and an altered tRNA profile but no changes in other Pol III transcripts. Thus, changes in the size and/or composition of the tRNA pool have a causal role in disease initiation. Our findings reveal different tissue- and brain region-specific sensitivities to a defect in Pol III transcription.

We are all different: Modeling key individual differences in physiological systems

Mathematical models of whole-body dynamics have advanced our understanding of human integrative systems that regulate physiological processes such as metabolism, temperature, and blood pressure. For most of these whole-body models, baseline parameters describe a 35-year-old young adult man who weighs 70 kg. As such, even among adults those models may not accurately represent half of the population (women), the older population, and those who weigh significantly more than 70 kg. Indeed, sex, age, and weight are known modulators of physiological function. To more accurately simulate a person who does not look like that “baseline person,” or to explain the mechanisms that yield the observed sex or age differences, these factors should be incorporated into mathematical models of physiological systems. Another key modulator is the time of day, because most physiological processes are regulated by the circadian clocks. Thus, ideally, mathematical models of integrative physiological systems should be specific to either a man or woman, of a certain age and weight, and a given time of day. We illustrate the importance of capturing these individual differences, using the blood pressure regulatory system as an example, and explain how that such models can be built.

Influence of X-ray spectrum and bowtie filter characterisation on the accuracy of Monte Carlo simulated organ doses: Validation in a whole-body CT scanning mode

PurposeFor patient-specific CT dosimetry, Monte Carlo dose simulations require an accurate description of the CT scanner. However, quantitative spectral information and information on the bowtie filter material and shape from the manufacturer is often not available. In this study, the influence of different X-ray spectra and bowtie filter characterisation methods on simulated CT organ doses is studied. MethodsUsing ImpactMC, organ doses of whole-body CTs were simulated in twenty adult whole-body voxel models, generated from PET/CT examinations previously conducted in these patients. Simulated CT organ doses based on the manufacturer X-ray spectra and bowtie filter data were compared with those obtained using alternative characterisation models, including spectrum generators and experimentally measured dose data. A total of four different X-ray spectra and one bowtie filter model were defined based on these data. ResultsFor all X-ray spectra and bowtie filter combinations, estimated CT organ doses are within 6% from those resulting from simulations with the CT characterisation models provided by the manufacturer. While varying the bowtie filter model results in CT organ dose differences smaller than 1%, dose differences up to 6% are observed when X-ray spectra are not based on the quantitative data from the manufacturer. ConclusionsEstimated organ doses slightly depend on the applied CT characterisation model. When manufacturer’s data are not available, half-value layer and dose measurements provide sufficient input to obtain equivalent X-ray spectra and bowtie filter profiles, respectively.

Study on Static Biomechanical Model of Whole Body Based on Virtual Human.

Material handling tasks often lead to skeletal injury of workers. The whole-body static biomechanical modeling method based on virtual humans is the theoretical basis for analyzing the human factor index in the lifting process. This paper focuses on the study of humans' body static biomechanical model for virtual human ergonomics analysis: First, the whole-body static biomechanical model is constructed, which calculates the biomechanical data such as force and moment, average strength, and maximum hand load at human joints. Secondly, the prototype model test system is developed, and the real experiment environment is set up with the inertial motion capture system. Finally, the model reliability verification experiment and application simulation experiment are designed. The comparison results with the industrial ergonomic software show that the model is consistent with the output of the industrial ergonomic software, which proves the reliability of the model. The simulation results show that under the same load, the maximum joint load and the maximum hand load are strongly related to the working posture, and the working posture should be adjusted to adapt to the load. Upright or bent legs have less influence on the maximum load capacity of the hand. Lower hand load capacity is due to forearm extension, and the upper arm extension greatly reduces the load capacity of the hand. Compared with a one-handed load, the two-handed load has a greater load capacity.

Maltodextrin-derived nanoparticles resensitize intracellular dormant Staphylococcus aureus to rifampicin

Intracellular bacteria are recognized as a crucial factor in the persistence and recurrence of infections. The efficacy of current antibiotic treatments faces substantial challenges due to the dormant state formation of intracellular bacteria. In this study, we devised a strategy aimed at reverting intracellular dormant bacteria to a metabolically active state, thereby increasing their vulnerability to antibiotics. We found that oligosaccharides, especially maltodextrin (MD), can be absorbed by dormant S. aureus, leading to their revival and restoration of sensitivity to rifampicin (Rif). We then synthesized a reactive oxygen species (ROS)-responsive MD-prodrug by covalently binding MD with 4-(hydroxymethyl) phenylboronic acid pinacol ester (MD-PBAP) and prepared a ROS-responsive nanoparticles (MDNP) using a nanoprecipitation and self-assembly method. Once internalized by host cells, MDNP was degraded to MD, reactivating dormant S. aureus, and enhancing their susceptibility to Rif. More importantly, MDNP treatment restored the sensitivity of intracellular persistent S. aureus to Rif in both a reservoir transfer model and whole-body infection model. Additionally, MDNP have demonstrated excellent biocompatibility in both in vitro and in vivo settings. These results offer a promising therapeutic avenue for managing persistent intracellular bacterial infections by reviving and resensitizing intracellular dormant bacteria to conventional antibiotics.

Hyperactive mTORC1/4EBP1 signaling dysregulates proteostasis and accelerates cardiac aging.

The mechanistic target of rapamycin complex 1 (mTORC1) has a major impact on aging by regulation of proteostasis. It is well established that mTORC1 signaling is hyperactivated with aging and age-related diseases. Previous studies have shown that partial inhibition of mTOR signaling by rapamycin reverses age-related deteriorations in cardiac function and structure in old mice. However, the downstream signaling pathways involved in this protection against cardiac aging have not been established. mTORC1 phosphorylates 4E-binding protein 1 (4EBP1) to promote the initiation of cap-dependent translation. The objective of this project is to examine the role of the mTORC1/4EBP1 axis in age-related cardiac dysfunction. We used a whole-body 4EBP1 KO mouse model, which mimics a hyperactive mTORC1/4EBP1/eIF4E axis, to investigate the effects of hyperactive mTORC1/4EBP1 axis in cardiac aging. Echocardiographic measurements of middle-aged 4EBP1 KO mice show impaired diastolic function and myocardial performance compared to age-matched WT mice and these parameters are at similar levels as old WT mice, suggesting that 4EBP1 KO mice experience accelerated cardiac aging. Old 4EBP1 KO mice show further decline in systolic and diastolic function compared to middle-aged counterparts and have worse systolic and diastolic function than age-matched WT mice. Gene expression levels of heart failure markers are not different between 4EBP1 KO and WT hearts. However, ribosomal biogenesis and protein ubiquitination are significantly increased in 4EBP1 KO hearts when compared to WT controls, suggesting dysregulated proteostasis in 4EBP1 KO hearts. Together, these results show that a hyperactive mTORC1/4EBP1 axis accelerates cardiac aging, potentially by dysregulating proteostasis.

Comparison of the difference in the anti-inflammatory activity of two different color types of Farfarae Flos based on in vitro, in vivo experiments and untargeted metabolomics.

Due to its remarkable anti-inflammatory pharmacological activity, Farfarae Flos has gained extensive usage in the treatment of various inflammatory diseases such as bronchitis, pneumonia, prostatitis and colitis. And Farfarae Flos come in two color types depending on the color of the flowers: yellowish-white (YW), and purplish-red (PR). However, the difference in anti-inflammatory activity and metabolic profiles between the two flower colors remains unexplored. This study aims to explore the difference in the anti-inflammatory potential between YW and PR variants of Farfarae Flos and unravel the mechanisms responsible for the observed differences in anti-inflammatory activity through an integrated approach encompassing untargeted metabolomics and in vivo/vitro experimental studies. Initially, we verified the contrasting effects of YW and PR on the inhibition of the inflammatory factors interleukin-6 (IL-6) and nitric oxide (NO) by utilizing an in vitro RAW 264.7 cell inflammation model. Subsequently, a comprehensive evaluation of the systemic inhibitory capacity of YW and PR on IL-6, Interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α) was conducted using a validated whole-body mouse model, followed by the analysis of inflammatory factors and histological examination of collected serum, liver, and spleen after 7days. Furthermore, non-targeted metabolomics profiling was employed to analyze the metabolite profiles of Farfarae Flos with different colors, and quantitative analysis was conducted to identify differential metabolites between YW and PR. The correlation between the anti-inflammatory activities of differentially accumulated metabolites (DAMs) and Farfarae Flos was investigated, resulting in the identification of 48 compounds exhibiting significant anti-inflammatory activity. Additionally, KEGG pathway enrichment analysis was performed to elucidate the underlying mechanisms. Our findings demonstrate that both YW and PR possess anti-inflammatory abilities, with PR exhibiting significantly superior efficacy. The integration of in vivo/vitro experiments and non-targeted metabolomics confirmed the exceptional anti-inflammatory potential of PR and solidified its classification as the "purplish-red better" of Farfarae Flos. This study provides valuable insights into the breeding and medical transformation of Farfarae Flos varieties, along with a scientific basis for the establishment of quality standards and the development of new drugs utilizing Farfarae Flos.

Volumetric versus Element-scaling Mass Estimation and Its Application to Permo-Triassic Tetrapods.

Size has an impact on various aspects of an animal's biology, including physiology, biomechanics, and ecology. Accurately and precisely estimating size, in particular body mass, is therefore a core objective of paleobiologists. Two approaches for estimating body mass are common: whole-body volumetric models and individual element-scaling (e.g., bones, teeth). The latter has been argued to be more accurate, while the former more precise. Here, we use minimum convex hulls (MCHs) to generate a predictive volumetric model for estimating body mass across a broad taxonomic and size range (127 g - 2735 kg). We compare our MCH model to stylopodial-scaling, incorporating data from the literature, and find that MCH body mass estimation is both more accurate and more precise than stylopodial estimation. An explanation for the difference between methods is that reptile and mammal stylopod circumference and length dimensions scale differentially (slope 1.179±0.102 vs. 1.038±0.031, respectively), such that reptiles have more robust bones for a given size. Consequently, a mammalian-weighted stylopodial-scaling sample overestimates the body mass of larger reptiles, and this error increases with size. We apply both estimation equations to a sample of 12 Permo-Triassic tetrapods and find that stylopodial-scaling consistently estimates a higher body mass than MCH estimation, due to even more robust bones in extinct species (slope=1.319±0.213). Finally, we take advantage of our MCH models to explore constraints regarding the position of the center of mass (CoM) and find that relative body proportions (i.e., skull:tail ratio) influence CoM position differently in mammals, crocodylians, and Permo-Triassic tetrapods. Further, we find that clade-specific body segment expansion factors do not affect group comparisons but may be important for individual specimens with rather disproportionate bodies (e.g., the small-headed and large-tailed Edaphosaurus). Our findings suggest that the whole-body volumetric approach is better suited for estimating body mass than element-scaling when anatomies are beyond the scope of the sample used to generate the scaling equations and provides added benefits such as the ability to measure inertial properties.

Spatial arrangement of the whiskers of harbor seals ( Phoca vitulina ) compared to whisker arrangements of house mice ( Mus musculus ) and brown rats ( Rattus norvegicus ).

We quantify the three-dimensional positions and orientations of the whiskers across the face of the harbor seal, and compare this geometry with the whisker arrays of rats and mice.

An Oxidative Stress Nano-Amplifier for Improved Tumor Elimination and Combined Immunotherapy.

Amplifying oxidative stress to disrupt intracellular redox homeostasis can accelerate tumor cell death. In this work, an oxidative stress amplifier (PP@T) is prepared for enhanced tumor oxidation therapy to reduce tumor growth and metastases. The nano-amplifier has been successfully constructed by embedding MTH1 inhibitor (TH588) in the PDA-coated porphyrin metal-organic framework PCN-224. The controllable-released TH588 is demonstrated from pores can hinder MTH1-mediated damage-repairing process by preventing the hydrolysis of 8-oxo-dG, thereby amplifying oxidative stress and exacerbating the oxidative DNA damage induced by the sonodynamic therapy of PP@T under ultrasound irradiation. Furthermore, PP@T can effectively induce immunogenic cell death to trigger systemic anti-tumor immune response. When administered in combination with immune checkpoint blockade, PP@T not only impedes the progression of the primary tumor but also achieves obvious antimetastasis in breast cancer murine models, including orthotopic and artificial whole-body metastasis models. Furthermore, the nanoplatform also provides photoacoustic imaging for in vivo treatment guidance. In conclusion, by amplifying oxidative stress and reactive oxygen species sensitized immunotherapy, this image-guided nanosystem shows potential for highly specific, effective combined therapy against tumor cells with negligible side-effects to normal cells which will provide a new insight for precise tumor treatment.

How rugose can you go? Spiny Agonidae armour decreases boundary layer separation

Abstract Armour has been present in vertebrates for millions of years and has evolved independently several times in the ray finned fishes. While armour is typically considered defensive, it is a multifunctional trait with many plausible alternative functions. We explore the hydrodynamic function of armour in nine species of poachers (Agonidae) by visualizing flow to assess drag reduction. We used microcomputed tomography scans to categorize armour morphology into morphotypes, and compared the rugosity (surface roughness) across these morphotypes. We then used digital particle image velocimetry to visualize boundary layer separation along 3D-printed whole-body models made using the scans. Poacher armour comprises eight rows that, some distance behind the second dorsal fin, merge into six. We found four morphotypes, with higher rugosity observed for the prominent spine morphotype and the lowest rugosity in the no-spine morphotype. Principal component analysis revealed that much of the variation in armour morphology is driven by plate shape, spine size, and protrusion. The boundary layer was retained longer in species with larger spines. Overall, the presence of a spine increased boundary layer retention, decreasing the drag, which is advantageous for these benthic fish.

In vivo dissection of the mouse tyrosine catabolic pathway with CRISPR-Cas9 identifies modifier genes affecting hereditary tyrosinemia type 1.

Hereditary tyrosinemia type 1 is an autosomal recessive disorder caused by mutations (pathogenic variants) in fumarylacetoacetate hydrolase, an enzyme involved in tyrosine degradation. Its loss results in the accumulation of toxic metabolites that mainly affect the liver and kidneys and can lead to severe liver disease and liver cancer. Tyrosinemia type 1 has a global prevalence of approximately 1 in 100,000 births but can reach up to 1 in 1,500 births in some regions of Québec, Canada. Mutating functionally related "modifier' genes (i.e. genes that, when mutated, affect the phenotypic impacts of mutations in other genes) is an emerging strategy for treating human genetic diseases. In vivo somatic genome editing in animal models of these diseases is a powerful means to identify modifier genes and fuel treatment development. In this study, we demonstrate that mutating additional enzymes in the tyrosine catabolic pathway through liver-specific genome editing can relieve or worsen the phenotypic severity of a murine model of tyrosinemia type 1. Neonatal gene delivery using recombinant adeno-associated viral vectors expressing Staphylococcus aureus Cas9 under the control of a liver-specific promoter led to efficient gene disruption and metabolic rewiring of the pathway, with systemic effects that were distinct from the phenotypes observed in whole-body knockout models. Our work illustrates the value of using in vivo genome editing in model organisms to study the direct effects of combining pathological mutations with modifier gene mutations in isogenic settings.

Modeling sex-specific whole-body metabolic responses to feeding and fasting

Men generally favor carbohydrate metabolism, while women lean towards lipid metabolism, resulting in significant sex-based differences in energy oxidation across various metabolic states such as fasting and feeding. These differences are influenced by body composition and inherent metabolic fluxes, including increased lipolysis rates in women. However, understanding how sex influences organ-specific metabolism and systemic manifestations remains incomplete. To address these gaps, we developed a sex-specific, whole-body metabolic model for feeding and fasting scenarios in healthy young adults. Our model integrates organ metabolism with whole-body responses to mixed meals, particularly high-carbohydrate and high-fat meals. Our predictions suggest that differences in liver and adipose tissue nutrient storage and oxidation patterns drive systemic metabolic disparities. We propose that sex differences in fasting hepatic glucose output may result from the different handling of free fatty acids, glycerol, and glycogen. We identified a metabolic pathway, possibly more prevalent in female livers, redirecting lipids towards carbohydrate metabolism to support hepatic glucose production. This mechanism is facilitated by the TG-FFA cycle between adipose tissue and the liver. Incorporating sex-specific data into multi-scale frameworks offers insights into how sex modulates human metabolism.

Understanding the metabolism of infants using whole-body metabolic models

A whole-body model is a computational representation of sex-specific and organ-resolved whole-body metabolism. In this issue of Cell Metabolism, Zaunseder etal. report whole-body models of infants that represent metabolic, physiological, energetic, and nutritional features, accurately simulating the growth of infants and providing foundations for personalized medicine for infants.

A whole-body mechanistic physiologically-based pharmacokinetic modeling of intravenous iron.

Iron is essential for every cell of the mammalian organism. Iron deficiency is a major public health issue worldwide. Intravenous (IV) iron therapy has been used to treat anemia. However, IV iron therapy is known far away from ideal because the quantitative relationship between the pharmacokinetics and biodistribution of IV iron under different iron statuses remains unclear. Patients are known to suffer adverse effects from excessive iron accumulation. Our objective was to develop a physiologically based pharmacokinetic (PBPK) model of iron in mice and validate its application for predicting iron disposition in rats and humans. Previously published data on iron were collected for constructing the PBPK model of iron in mice, and then extrapolated to rats and humans based on physiologically and chemically specific parameters relevant to each species. The PBPK model characterized the distribution of iron in mice successfully. The model based on extrapolation to rats accurately simulated the ferric carboxymaltose (FCM) PK profiles in rat tissues. Similarly, the observed and simulated serum PK of FCM in humans were in reasonable agreement. This mechanistic whole-body PBPK model is useful for understanding and predicting iron effects on different species. It also establishes a foundation for future research that incorporates iron kinetics and biodistribution, along with related clinical experiments. This approach could lead to the development of effective and personalized iron deficiency anemia treatments.

Molecular basis of neurodegeneration in a mouse model of Polr3-related disease.

Pathogenic variants in subunits of RNA polymerase (Pol) III cause a spectrum of Polr3-related neurodegenerative diseases including 4H leukodystrophy. Disease onset occurs from infancy to early adulthood and is associated with a variable range and severity of neurological and non-neurological features. The molecular basis of Polr3-related disease pathogenesis is unknown. We developed a postnatal whole-body mouse model expressing pathogenic Polr3a mutations to examine the molecular mechanisms by which reduced Pol III transcription results primarily in central nervous system phenotypes. Polr3a mutant mice exhibit behavioral deficits, cerebral pathology and exocrine pancreatic atrophy. Transcriptome and immunohistochemistry analyses of cerebra during disease progression show a reduction in most Pol III transcripts, induction of innate immune and integrated stress responses and cell type-specific gene expression changes reflecting neuron and oligodendrocyte loss and microglial activation. Earlier in the disease when integrated stress and innate immune responses are minimally induced, mature tRNA sequencing revealed a global reduction in tRNA levels and an altered tRNA profile but no changes in other Pol III transcripts. Thus, changes in the size and/or composition of the tRNA pool have a causal role in disease initiation. Our findings reveal different tissue- and brain region-specific sensitivities to a defect in Pol III transcription.