White matter fiber integrity and structural brain network topology: implications for balance function in postischemic stroke patients.

Previous studies have suggested that ischemic stroke can result in white matter fiber injury and modifications in the structural brain network. However, the relationship with balance function scores remains insufficiently explored. Therefore, this study aims to explore the alterations in the microstructural properties of brain white matter and the topological characteristics of the structural brain network in postischemic stroke patients and their potential correlations with balance function. We enrolled 21 postischemic stroke patients and 21 age, sex, and education-matched healthy controls (HC). All participants underwent balance function assessment and brain diffusion tensor imaging. Tract-based spatial statistics (TBSS) were used to compare the fractional anisotropy, mean diffusivity, axial diffusivity, and radial diffusivity of white matter fibers between the two groups. The white matter structural brain network was constructed based on the automated anatomical labeling atlas, and we conducted a graph theory-based analysis of its topological properties, including global network properties and local node properties. Additionally, the correlation between the significant structural differences and balance function score was analyzed. The TBSS results showed that in comparison to the HC, postischemic stroke patients exhibited extensive damage to their whole-brain white matter fiber tracts (P < 0.05). Graph theory analysis showed that in comparison to the HC, postischemic stroke patients exhibited statistically significant reductions in the values of global efficiency, local efficiency, and clustering coefficient, as well as an increase in characteristic path length (P < 0.05). In addition, the degree centrality and nodal efficiency of some nodes in postischemic stroke patients were significantly reduced (P < 0.05). The white matter fibers of the entire brain in postischemic stroke patients are extensively damaged, and the topological properties of the structural brain network are altered, which are closely related to balance function. This study is helpful in further understanding the neural mechanism of balance function after ischemic stroke from the white matter fiber and structural brain network topological properties.

In the Murine and Bovine Maternal Mammary Gland Signal Transducer and Activator of Transcription 3 is Activated in Clusters of Epithelial Cells around the Day of Birth.

Signal transducers and activators of transcription (STAT) proteins regulate mammary development. Here we investigate the expression of phosphorylated STAT3 (pSTAT3) in the mouse and cow around the day of birth. We present localised colocation analysis, applicable to other mammary studies requiring identification of spatially congregated events. We demonstrate that pSTAT3-positive events are multifocally clustered in a non-random and statistically significant fashion. Arginase-1 expressing cells, consistent with macrophages, exhibit distinct clustering within the periparturient mammary gland. These findings represent a new facet of mammary STAT3 biology, and point to the presence of mammary sub-microenvironments.

Assessing the tumor immune landscape across multiple spatial scales to differentiate immunotherapy response in metastatic non-small cell lung cancer.

While immune checkpoint inhibitor-based (ICI) therapy has shown promising results in non-small cell lung cancer (NSCLC) patients with high programmed death ligand 1 (PD-L1) expression, not all patients respond to therapy. The tumor microenvironment (TME) is complex and heterogeneous, making it challenging to understand the key agents and features which influence response to therapies. In this study, we leverage multiplex fluorescent immunohistochemistry (mfIHC) to quantitatively assess interactions between tumor and immune cells in an effort to identify patterns occurring at multiple spatial levels of the TME. To do so, we introduce several computational methods novel to a dataset of 1,269 mfIHC images from a cohort of 52 patients with metastatic NSCLC. With the spatial G-cross function, we quantify the degree of cell interaction at an entire image level, where we see significantly increased activity of cytotoxic T-cells (CTLs) and helper T-cells (HTLs) with epithelial tumor cells (ECs) in responders to ICI (p = .022 and p < .001, respectively), and decreased activity of T-regulatory cells (Tregs) with ECs compared to non-responders (p = .010). By leveraging spatial overlap methods, we define tumor subregions (which we call the tumor "periphery", "edge" and "center") and discover more localized immune-immune interactions influencing positive response, including those between CTLs and HTLs with antigen presenting cells (APCs) in these subregions specifically. Lastly, we trained an interpretable deep learning model which identified key cellular regions of interest that most influenced response classification (AUC = 0.71±0.02). Assessing spatial interactions within these subregions further revealed new insights not significant at the whole image level, particularly the elevated association of APCs and Tregs with one another in responder groups (p = 0.024). Altogether, we demonstrate that elucidating patterns of cell composition and interplay across multiple levels of spatial analyses can improve our understanding of the TME and better differentiate patient responses to immunotherapy.

An introduction to bayesian spatial smoothing methods for disease mapping: modeling county firearm suicide mortality rates.

This article introduces bayesian spatial smoothing models for disease mapping-a specific application of small area estimation where the full universe of data is known-to a wider audience of public health professionals using firearm suicide as a motivating example. Besag, York, and Mollié (BYM) Poisson spatial and space-time smoothing models were fitted to firearm suicide counts for the years 2014-2018. County raw death rates in 2018 ranged from 0 to 24.81 deaths per 10 000 people. However, the highest mortality rate was highly unstable, based on only 2 deaths in a population of approximately 800, and 80.5% of contiguous US counties experienced fewer than 10 firearm suicide deaths and were thus suppressed. Spatially smoothed county firearm suicide mortality estimates ranged from 0.06 to 4.05 deaths per 10 000 people and could be reported for all counties. The space-time smoothing model produced similar estimates with narrower credible intervals as it allowed counties to gain precision from adjacent neighbors and their own counts in adjacent years. bayesian spatial smoothing methods are a useful tool for evaluating spatial health disparities in small geographies where small numbers can result in highly variable rate estimates, and new estimation techniques in R software have made fitting these models more accessible to researchers.

Alleviating spatial confounding in frailty models.

The confounding between fixed effects and (spatial) random effects in a regression setup is termed spatial confounding. This topic continues to gain attention and has been studied extensively in recent years, given that failure to account for this may lead to a suboptimal inference. To mitigate this, a variety of projection-based approaches under the class of restricted spatial models are available in the context of generalized linear mixed models. However, these projection approaches cannot be directly extended to the spatial survival context via frailty models due to dimension incompatibility between the fixed and spatial random effects. In this work, we introduce a two-step approach to handle this, which involves (i) projecting the design matrix to the dimension of the spatial effect (via dimension reduction) and (ii) assuring that the random effect is orthogonal to this new design matrix (confounding alleviation). Under a fully Bayesian paradigm, we conduct fast estimation and inference using integrated nested Laplace approximation. Both simulation studies and application to a motivating data evaluating respiratory cancer survival in the US state of California reveal the advantages of our proposal in terms of model performance and confounding alleviation, compared to alternatives.

Microstructural white matter abnormalities in overactive bladder syndrome evaluation with diffusion kurtosis imaging tract-based spatial statistics analysis.

PURPOSE: This prospective study aimed to explore the microstructural alterations of the white matter in overactive bladder syndrome (OAB) using the Tract-based Spatial Statistics (TBSS) method of diffusion kurtosis imaging (DKI). METHODS: A total of 30 patients were enrolled and compared with 30 controls. White matter (WM) status was assessed using tract-based spatial statistics for DKI. The differences in DKI-derived parameters, including kurtosis fractional anisotropy (KFA), fractional anisotropy (FA), mean kurtosis (MK), mean diffusivity (MD), radial kurtosis (RK), axial kurtosis (AK), axial diffusivity (AD), and radial diffusivity (RD), were compared between the two groups using the TBSS method. The correlation between the altered DKI-derived parameters and the (OABSS) scores was analyzed. A receiver operating characteristic curve (ROC) was used to evaluate the diagnostic performance of different white matter parameters. RESULTS: As a result, compared with the HC group, the KFA, and FA values decreased significantly in the OAB group. Compared with the HC group, the MK and MD values increased significantly in the OAB group. The KFA values of the genu of corpus callosum (GCC) were significantly correlated with the OABSS scores (r = - 0.509; p = 0.004). The FA values of anterior corona radiata (ACR) were significantly correlated with OABSS scores (r = - 0.447; p = 0.013). The area under the ROC curve (AUC) for the genu of corpus callosum KFA values was higher than FA for the diagnosis of OAB patients. CONCLUSION: DKI is a promising approach to the investigation of the pathophysiology of OAB and a potential biomarker for clinical diagnosis of OAB.

Visibility graph-based covariance functions for scalable spatial analysis in non-convex partially Euclidean domains.

We present a new method for constructing valid covariance functions of Gaussian processes for spatial analysis in irregular, non-convex domains such as bodies of water. Standard covariance functions based on geodesic distances are not guaranteed to be positive definite on such domains, while existing non-Euclidean approaches fail to respect the partially Euclidean nature of these domains where the geodesic distance agrees with the Euclidean distances for some pairs of points. Using a visibility graph on the domain, we propose a class of covariance functions that preserve Euclidean-based covariances between points that are connected in the domain while incorporating the non-convex geometry of the domain via conditional independence relationships. We show that the proposed method preserves the partially Euclidean nature of the intrinsic geometry on the domain while maintaining validity (positive definiteness) and marginal stationarity of the covariance function over the entire parameter space, properties which are not always fulfilled by existing approaches to construct covariance functions on non-convex domains. We provide useful approximations to improve computational efficiency, resulting in a scalable algorithm. We compare the performance of our method with those of competing state-of-the-art methods using simulation studies on synthetic non-convex domains. The method is applied to data regarding acidity levels in the Chesapeake Bay, showing its potential for ecological monitoring in real-world spatial applications on irregular domains.

Latent Archetypes of the Spatial Patterns of Cancer.

The cancer atlas edited by several countries is the main resource for the analysis of the geographic variation of cancer risk. Correlating the observed spatial patterns with known or hypothesized risk factors is time-consuming work for epidemiologists who need to deal with each cancer separately, breaking down the patterns according to sex and race. The recent literature has proposed to study more than one cancer simultaneously looking for common spatial risk factors. However, this previous work has two constraints: they consider only a very small (2-4) number of cancers previously known to share risk factors. In this article, we propose an exploratory method to search for latent spatial risk factors of a large number of supposedly unrelated cancers. The method is based on the singular value decomposition and nonnegative matrix factorization, it is computationally efficient, scaling easily with the number of regions and cancers. We carried out a simulation study to evaluate the method's performance and apply it to cancer atlas from the USA, England, France, Australia, Spain, and Brazil. We conclude that with very few latent maps, which can represent a reduction of up to 90% of atlas maps, most of the spatial variability is conserved. By concentrating on the epidemiological analysis of these few latent maps a substantial amount of work is saved and, at the same time, high-level explanations affecting many cancers simultaneously can be reached.

Bayesian modeling of spatial ordinal data from health surveys.

Health surveys allow exploring health indicators that are of great value from a public health point of view and that cannot normally be studied from regular health registries. These indicators are usually coded as ordinal variables and may depend on covariates associated with individuals. In this article, we propose a Bayesian individual-level model for small-area estimation of survey-based health indicators. A categorical likelihood is used at the first level of the model hierarchy to describe the ordinal data, and spatial dependence among small areas is taken into account by using a conditional autoregressive distribution. Post-stratification of the results of the proposed individual-level model allows extrapolating the results to any administrative areal division, even for small areas. We apply this methodology to describe the geographical distribution of a self-perceived health indicator from the Health Survey of the Region of Valencia (Spain) for the year 2016.

Modeling variation in mixture effects over space with a Bayesian spatially varying mixture model.

Mixture analysis is an emerging statistical tool in epidemiological research that seeks to estimate the health effects associated with mixtures of several exposures. This approach acknowledges that individuals experience many simultaneous exposures and it can estimate the relative importance of components in the mixture. Health effects due to mixtures may vary over space driven by to political, demographic, environmental, or other differences. In such cases, estimating a global mixture effect without accounting for spatial variation would induce bias in effect estimates and potentially lower statistical power. To date, no methods have been developed to estimate spatially varying chemical mixture effects. We developed a Bayesian spatially varying mixture model that estimates spatially varying mixture effects and the importance weights of components in the mixture, while adjusting for covariates. We demonstrate the efficacy of the model through a simulation study that varies the number of mixtures (one and two) and spatial pattern (global, one-dimensional, radial) and magnitude of mixture effects, showing that the model is able to accurately reproduce the spatial pattern of mixture effects across a diverse set of scenarios. Finally, we apply our model to a multi-center case-control study of non-Hodgkin lymphoma (NHL) in Detroit, Iowa, Los Angeles, and Seattle. We identify significant spatially varying positive and inverse associations with NHL for two mixtures of pesticides in Iowa and do not find strong spatial effects at the other three centers. In conclusion, the Bayesian spatially varying mixture model represents a novel method for modeling spatial variation in mixture effects.

Spatial confidence regions for combinations of excursion sets in image analysis.

The analysis of excursion sets in imaging data is essential to a wide range of scientific disciplines such as neuroimaging, climatology, and cosmology. Despite growing literature, there is little published concerning the comparison of processes that have been sampled across the same spatial region but which reflect different study conditions. Given a set of asymptotically Gaussian random fields, each corresponding to a sample acquired for a different study condition, this work aims to provide confidence statements about the intersection, or union, of the excursion sets across all fields. Such spatial regions are of natural interest as they directly correspond to the questions 'Where do all random fields exceed a predetermined threshold?', or 'Where does at least one random field exceed a predetermined threshold?'. To assess the degree of spatial variability present, our method provides, with a desired confidence, subsets and supersets of spatial regions defined by logical conjunctions (i.e. set intersections) or disjunctions (i.e. set unions), without any assumption on the dependence between the different fields. The method is verified by extensive simulations and demonstrated using task-fMRI data to identify brain regions with activation common to four variants of a working memory task.

Spatial analyses of immune cell infiltration in cancer: current methods and future directions: A report of the International Immuno-Oncology Biomarker Working Group on Breast Cancer.

Modern histologic imaging platforms coupled with machine learning methods have provided new opportunities to map the spatial distribution of immune cells in the tumor microenvironment. However, there exists no standardized method for describing or analyzing spatial immune cell data, and most reported spatial analyses are rudimentary. In this review, we provide an overview of two approaches for reporting and analyzing spatial data (raster versus vector-based). We then provide a compendium of spatial immune cell metrics that have been reported in the literature, summarizing prognostic associations in the context of a variety of cancers. We conclude by discussing two well-described clinical biomarkers, the breast cancer stromal tumor infiltrating lymphocytes score and the colon cancer Immunoscore, and describe investigative opportunities to improve clinical utility of these spatial biomarkers. © 2023 The Pathological Society of Great Britain and Ireland.

White and Gray Matter Abnormalities in Young Adult Females with Dependent Personality Disorder: A Diffusion-Tensor Imaging and Voxel-Based Morphometry Study.

We applied diffusion-tensor imaging (DTI) including measurements of fractional anisotropy (FA), a parameter of neuronal fiber integrity, mean diffusivity (MD), a parameter of brain tissue integrity, as well as voxel-based morphometry (VBM), a measure of gray and white matter volume, to provide a basis to improve our understanding of the neurobiological basis of dependent personality disorder (DPD). DTI was performed on young girls with DPD (N = 17) and young female healthy controls (N = 17). Tract-based spatial statistics (TBSS) were used to examine microstructural characteristics. Gray matter volume differences between the two groups were investigated using voxel-based morphometry (VBM). The Pearson correlation analysis was utilized to examine the relationship between distinct brain areas of white matter and gray matter and the Dy score on the MMPI. The DPD had significantly higher fractional anisotropy (FA) values than the HC group in the right retrolenticular part of the internal capsule, right external capsule, the corpus callosum, right posterior thalamic radiation (include optic radiation), right cerebral peduncle (p < 0.05), which was strongly positively correlated with the Dy score of MMPI. The volume of gray matter in the right postcentral gyrus and left cuneus in DPD was significantly increased (p < 0.05), which was strongly positively correlated with the Dy score of MMPI (r1,2= 0.467,0.353; p1,2 = 0.005,0.04). Our results provide new insights into the changes in the brain structure in DPD, which suggests that alterations in the brain structure might implicate the pathophysiology of DPD. Possible visual and somatosensory association with motor nerve circuits in DPD.

Evaluation of microstructural changes in the brain in transfusion dependent thalassemia patients with advanced magnetic resonance imaging techniques.

PURPOSE: Transfusion-dependent thalassemia (TDT) is associated with iron accumulation in the body and an increased tendency for thrombosis. With the increased life expectancy in these patients, the detection of neurocognitive complications has gained importance. This study investigates the microstructural changes in TDT patients using advanced diffusion MRI techniques and their relationship with laboratory parameters. METHODS: The study included 14 TDT patients and 14 control subjects. Tract-based spatial statistics (TBSS) were used to examine differences in DTI parameters such as fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) in thalassemia patients using multi-shell DWI images. The mean kurtosis (MK) difference was investigated using diffusion kurtosis imaging. Fiber density (FD), fiber cross-section (FC), and fiber density and cross-section (FDC) differences were examined using fixel-based analysis. In the patient group, correlative tractography was used to investigate the relationship between DTI parameters and platelet (PLT) and ferritin levels. RESULTS: Increase in RD and MD was observed, particularly in the white matter tracts of the corona radiata in patient group. Additionally, an increase in AD was detected in a limited area. Correlative tractography in thalasemia patients showed a positive correlation between increases in RD, MD, and AD with PLT and ferritin. Fixel-based analysis demonstrated a dispersed distribution in white matter fibers, with a more pronounced decrease in FD, FC, and FDC in the internal capsule. CONCLUSION: There is widespread involvement in the white matter and fiber tracts in thalassemia patients, which is highly correlated with thrombotic parameters.

Combined diffusion tensor imaging and quantitative susceptibility mapping to characterize normal-appearing white matter in self-limited epilepsy with centrotemporal spikes.

PURPOSE: In brain development, Myelination is the characteristic feature of white matter maturation, which plays an important role in efficient information transmitting. The white matter abnormality has been reported to be associated with self-limited epilepsy with centrotemporal spikes (SeLECTS). This study aimed to detect the altered white matter region in the SeLECTS patients by the combination of diffusion tensor imaging (DTI) and quantitative susceptibility mapping (QSM) technique. METHODS: 27 children with SeLECTS and 23 age- and gender-matched healthy children were enrolled. All participants were scanned with 3.0-T MRI to acquire the structure, diffusion and susceptibility-weighted data. The susceptibility and diffusion weighted data were processed to obtain quantitative susceptibility map and fraction anisotropy (FA) map. Then voxel-wise tract-based spatial statistics (TBSS) were used to analyze quantitative susceptibility and FA data. RESULTS: Both DTI and QSM revealed extensive white matter alterations in the frontal, parietal, and temporal lobes in SeLECTS patients. The overlapped region of DTI and QSM analyses was located in the fiber tracts of the corona radiata. The FA values in this overlapped region were negatively correlated with the magnetic susceptibility values. CONCLUSION: Our results suggest that TBSS-based QSM can be employed as a novel approach for characterizing alterations in white matter in SeLECTS. And the combination of QSM and DTI can provide a more comprehensive evaluation of white matter integrity by utilizing different biophysical features.

Facial emotion recognition function and white matter microstructural alterations in drug-naive, comorbidity-free autism.

Individuals with autism spectrum disorder have deficits in facial emotion recognition and white matter microstructural alterations. Nonetheless, most previous studies were confounded by different variables, such as psychiatric comorbidities and psychotropic medications used by ASD participants. Also, it remains unclear how exactly FER deficits are related to white matter microstructural alterations in ASD. Accordingly, we aimed to investigate the FER functions, white matter microstructure, and their relationship in drug-naive and comorbidity-free ASD individuals. 59 ASD individuals and 59 typically developed individuals were included, where 46 ASD and 50 TD individuals completed FER tasks. Covariance analysis showed scores were lower in both basic and complex FER tasks in the ASD group. Tract-Based Spatial Statistics showed FA values in widespread white matter fibers were lower in the ASD group than in the TD group, including forceps major and forceps minor of the corpus callosum, anterior thalamic radiation, corticospinal tract, cingulum, inferior frontal-occipital fasciculus, inferior longitudinal fasciculus, superior longitudinal fasciculus. Moreover, in the TD group but not the ASD group, the performance in the complex FER task was negatively correlated with the FA value in some white matter fibers, including forceps major of the corpus callosum, ATR, CT, cingulum, IFOF, ILF, SLF. Our study suggests children with ASD may experience deficits in facial emotion recognition and exhibit alterations in white matter microstructure. More importantly, our study indicates that white matter microstructural alterations may be involved in FER deficits in children with ASD.

Plasma Apo-E mediated corticospinal tract abnormalities and suicidality in patients with major depressive disorder.

This study aims to explore the link between Apo-E, brain white matter, and suicide in patients with major depressive disorder (MDD) to investigate the potential neuroimmune mechanisms of Apo-E that may lead to suicide. Thirty-nine patients with MDD (22 patients with suicidality) and 57 age, gender, and education-matched healthy controls participated in this study, provided plasma Apo-E samples, and underwent diffusion tensor imaging scans. Plasma Apo-E levels and white matter microstructure were analyzed among the MDD with suicidality, MDD without suicidality, and HC groups using analysis of variance with post hoc Bonferroni correction and tract-based spatial statistics (TBSS) with threshold-free cluster enhancement correction. Mediation analysis investigated the relationship between Apo-E, brain white matter, and suicidality in MDD. The MDD with suicidality subgroup had higher depressive and suicide scores, longer disease course, and lower plasma Apo-E levels than MDD without suicidality. TBSS revealed that the MDD non-suicide subgroup showed significantly increased mean diffusivity in the left corticospinal tract and body of the left corpus callosum, as well as increased axial diffusivity in the left anterior corona radiata and the right posterior thalamic radiation compared to the suicidal MDD group. The main finding was that the increased MD of the left corticospinal tract contributed to the elevated suicide score, with Apo-E mediating the effect. Preliminary result that Apo-E's mediating role between the left corticospinal tract and the suicide factor suggests the neuroimmune mechanism of suicide in MDD. The study was registered on ClinicalTrials.gov (NCT03790085).

Altered cortical microstructure in preterm infants at term-equivalent age relative to term-born neonates.

Preterm (PT) birth is a potential factor for abnormal brain development. Although various alterations of cortical structure and functional connectivity in preterm infants have been reported, the underlying microstructural foundation is still undetected thoroughly in PT infants relative to full-term (FT) neonates. To detect the very early cortical microstructural alteration noninvasively with advanced neurite orientation dispersion and density imaging (NODDI) on a whole-brain basis, we used multi-shell diffusion MRI of healthy newborns selected from the Developing Human Connectome Project. 73 PT infants and 69 FT neonates scanned at term-equivalent age were included in this study. By extracting the core voxels of gray matter (GM) using GM-based spatial statistics (GBSS), we found that comparing to FT neonates, infants born preterm showed extensive lower neurite density in both primary and higher-order association cortices (FWE corrected, P < 0.025). Higher orientation dispersion was only found in very preterm subgroup in the orbitofrontal cortex, fronto-insular cortex, entorhinal cortex, a portion of posterior cingular gyrus, and medial parieto-occipital cortex. This study provided new insights into exploring structural MR for functional and behavioral variations in preterm population, and these findings may have marked clinical importance, particularly in the guidance of ameliorating the development of premature brain.

A New Method for Low Density Distribution Modeling and Near Threatened Species: The Study Case of Plectrohyla Guatemalensis.

We introduce a model that can be used for the description of the distribution of species when there is scarcity of data, based on our previous work (Ballesteros et al. J Math Biol 85(4):31, 2022). We address challenges in modeling species that are seldom observed in nature, for example species included in The International Union for Conservation of Nature's Red List of Threatened Species (IUCN 2023). We introduce a general method and test it using a case study of a near threatened species of amphibians called Plectrohyla Guatemalensis (see IUCN 2023) in a region of the UNESCO natural reserve "Tacaná Volcano", in the border between Mexico and Guatemala. Since threatened species are difficult to find in nature, collected data can be extremely reduced. This produces a mathematical problem in the sense that the usual modeling in terms of Markov random fields representing individuals associated to locations in a grid generates artificial clusters around the observations, which are unreasonable. We propose a different approach in which our random variables describe yearly averages of expectation values of the number of individuals instead of individuals (and they take values on a compact interval). Our approach takes advantage of intuitive insights from environmental properties: in nature individuals are attracted or repulsed by specific features (Ballesteros et al. J Math Biol 85(4):31, 2022). Drawing inspiration from quantum mechanics, we incorporate quantum Hamiltonians into classical statistical mechanics (i.e. Gibbs measures or Markov random fields). The equilibrium between spreading and attractive/repulsive forces governs the behavior of the species, expressed through a global control problem involving an energy operator.

Risk factors for persistent fatal opioid-involved overdose clusters in Massachusetts 2011-2021: a spatial statistical analysis with socio-economic, accessibility, and prescription factors.

BACKGROUND: Fatal opioid-involved overdose rates increased precipitously from 5.0 per 100,000 population to 33.5 in Massachusetts between 1999 and 2022. METHODS: We used spatial rate smoothing techniques to identify persistent opioid overdose-involved fatality clusters at the ZIP Code Tabulation Area (ZCTA) level. Rate smoothing techniques were employed to identify locations of high fatal opioid overdose rates where population counts were low. In Massachusetts, this included areas with both sparse data and low population density. We used Local Indicators of Spatial Association (LISA) cluster analyses with the raw incidence rates, and the Empirical Bayes smoothed rates to identify clusters from 2011 to 2021. We also estimated Empirical Bayes LISA cluster estimates to identify clusters during the same period. We constructed measures of the socio-built environment and potentially inappropriate prescribing using principal components analysis. The resulting measures were used as covariates in Conditional Autoregressive Bayesian models that acknowledge spatial autocorrelation to predict both, if a ZCTA was part of an opioid-involved cluster for fatal overdose rates, as well as the number of times that it was part of a cluster of high incidence rates. RESULTS: LISA clusters for smoothed data were able to identify whether a ZCTA was part of a opioid involved fatality incidence cluster earlier in the study period, when compared to LISA clusters based on raw rates. PCA helped in identifying unique socio-environmental factors, such as minoritized populations and poverty, potentially inappropriate prescribing, access to amenities, and rurality by combining socioeconomic, built environment and prescription variables that were highly correlated with each other. In all models except for those that used raw rates to estimate whether a ZCTA was part of a high fatality cluster, opioid overdose fatality clusters in Massachusetts had high percentages of Black and Hispanic residents, and households experiencing poverty. The models that were fitted on Empirical Bayes LISA identified this phenomenon earlier in the study period than the raw rate LISA. However, all the models identified minoritized populations and poverty as significant factors in predicting the persistence of a ZCTA being part of a high opioid overdose cluster during this time period. CONCLUSION: Conducting spatially robust analyses may help inform policies to identify community-level risks for opioid-involved overdose deaths sooner than depending on raw incidence rates alone. The results can help inform policy makers and planners about locations of persistent risk.