Spatial Omics: Clustering and Integration

Yingshan Qiu Chair
Auburn University
 
Tuesday, Aug 4: 10:30 AM - 12:20 PM
6426 
Contributed Papers 
Thomas M. Menino Convention & Exhibition Center 
Room: CC-254A 

Main Sponsor

Section on Statistics in Genomics and Genetics

Presentations

Cell class distributional learning for low-resolution spatially transcriptomic data

Recent advancements in technology have made it possible to profile spatial transcriptomics (ST), which captures genome-wide gene expression while maintaining the spatial arrangement of cells within tissues. This progress has led to a better understanding of cell communication and tissue organization. However, current techniques face a trade-off between experimental throughput and spatial resolution. Sequencing-based approaches tend to prioritize higher throughput, which results in lower resolution, producing multicellular pixel data. This type of data necessitates innovative computational methods to disentangle cell classes within a pixel and address potential confounding issues. In this study, we are interested in learning the cell class-specific information from low-resolution ST data, where each pixel contains a mixture of several different cell classes. We develop a framework leveraging topic modeling, gene network, and machine-learning to infer the distribution of cell classes across regions and genes. 

Keywords

Spatial transcriptomic data

Low-resolution ST data

Topic modeling

Gene network

Cell class distribution

Cell-cell communication 

Speaker

Wooyoung Kim, Washington State University

Co-Author

Yuan Wang, Washington State University

Controlling False Discoveries after Clustering via Data Splitting for Spatial Marker Detection

Spatial omics technologies are transforming biomedical research by enabling genome-wide measurement of molecular activity while preserving the spatial context within tissues. These advances create unprecedented opportunities to uncover cell–cell interactions, tissue organization, and disease mechanisms. A crucial step in realizing this potential is identifying spatial domain markers, which are essential for defining tissue architecture and understanding disease progression. However, using the same data for both clustering and marker detection creates the problem of "double dipping", which can lead to inflated false discoveries, particularly when domain boundaries are poorly defined. To address this challenge, we develop SpaDS and SpaMDS, data splitting based approaches for differential expression testing after clustering for spatial omics, enabling robust spatial domain marker discovery with controlled false discovery rate and high power. Through extensive simulations and analyses of spatial omics datasets, we demonstrate that the data-splitting methods are easy to implement, adaptable to existing spatial omics data analysis pipelines, and often outperform other approaches. 

Keywords

Statistical genomics

Spatial transcriptomics

data splitting

FDR control 

Speaker

Yingxin Lin, The Chinese University of Hong Kong

Co-Author(s)

Lijun Wang
Hongyu Zhao, Yale University

Manifold‑Informed Gene‑module Extraction for Disentangling Simultaneous Dynamics in scRNA‑Seq

We introduce MIRAGE, a statistical framework using cell-cell manifolds to test whether gene sets encode the same or distinct cellular dynamics. MIRAGE constructs manifolds for many gene sets, quantifies their geometric similarity, and uses hypothesis testing to merge only sets sharing common cell–cell geometry, identifying independent or co-occurring cellular dynamics. Applied to mouse pancreatic endocrinogenesis, MIRAGE separates developmental and cell-cycle programs, and in fibroblast reprogramming datasets, yields reproducible gene modules tracking shared intermediate states. In human Alzheimer's disease microglia, where heterogeneous stimuli produce overlapping homeostatic, inflammatory, and metabolic states, MIRAGE identifies one module capturing a within-donor continuum in both AD and non-AD brains, and a second branching module with one arm enriched for hypoxia and lysosomal stress genes, preferentially occupied by donors with elevated amyloid/tau burden and cognitive impairment. MIRAGE offers a principled approach for decomposing single-cell data into pathway-level manifolds, clarifying how co-occurring cellular programs contribute to disease-relevant heterogeneity. 

Keywords

Gene clustering

Geometric structure

Nearest neighbor graph

Multivariate hypothesis testing 

Speaker

Zhaoheng Li

Co-Author

Kevin Lin, University of Washington

POLYspace reveals domain-specific cellular neighborhoods through topology-aware spatial analysis

Identifying cellular neighborhoods is essential for understanding cell–cell interactions in a spatial context. However, existing approaches often overlook the complexity of multicellular interactions and the organization of spatial domains. We present POLYspace, a general and efficient framework for discovering and analyzing cellular neighborhoods of arbitrary topology while accounting for spatial domains. POLYspace formulates neighborhood identification as a subgraph searching problem and leverages C3G, a fast graph canonization algorithm we developed, to achieve scalability. Applied to one in-house dataset and three publicly available datasets spanning diverse platforms and tissues, POLYspace uncovers domain-specific cellular neighborhoods that are not captured by existing methods. These neighborhoods reveal key biological mechanisms and improve phenotype prediction. 

Keywords

Spatial gene profiling

Graph theory

Graph canonization

Statistical genomics

Subgraph mining

Cellular microenvironments 

Speaker

Huimin Wang, Duke University School of Medicine Dept. of Biostatistics & Bioinformation

Co-Author(s)

Roger McLendon, Department of Pathology, Duke University School of Medicine
Simon Gregory, Department of Neurosurgery, Duke University School of Medicine
Zhicheng Ji, Duke University

Scale-Invariant Joint Clustering and Deconvolution of Spatial Transcriptomics

We propose a statistical framework for simultaneous clustering and deconvolution of spatial transcriptomics (ST) that is reference-free. Our method jointly estimates the reference cell-type-specific gene expression profiles and the cluster assignments and their cell-type composition of ST spots through solving an well-defined optimization problem. We introduce a regularization term that makes the problem scale invariant which separates compositional signatures from library‑size effects and enabling clustering that groups spots by true compositional similarity even when counts differ. Furthermore, we leverage spatial information to encourage neighboring spots to have similar cell-type compositions. We develop a computationally efficient algorithm to solve the optimization problem and establish theoretical properties of our estimator. Through extensive simulations and applications to real ST data, we demonstrate that our method outperforms existing reference-free methods and is able to uncover biologically meaningful clusters and accurately estimate cell-type compositions without relying on external single-cell references. 

Keywords

Clustering

Spatial Transcriptomics

Scale invariant

Post selection inference

Optimization

Reference‑free deconvolution 

Speaker

Hyun Jung Koo, School of Statistics, University of Minnesota - Twin Cities

Co-Author

Aaron Molstad, University of Minnesota

SpatialCCA: Spatially aware method for cross-tissue integration in Spatial Transcriptomics

Spatial transcriptomics (ST) enables genome-wide expression profiling with preserved spatial context. However, integrating multiple tissue sections remains challenging due to the lack of direct spatial correspondence across different samples. We present SpatialCCA, a method for joint dimension reduction of two ST tissue sections that embeds them into a shared latent space while incorporating spatial information. SpatialCCA extends canonical correlation analysis through a kernel-based spatial regularization and models tissue-specific variation to mitigate batch effects. The resulting embedding captures both cross-section transcriptional similarity and within-tissue spatial organization. Using simulation generated by randomly partitioning real ST tissue data into two sets of spatial spots, we show that SpatialCCA yields embeddings with stronger correlation with the original coordinates compared to embeddings derived from gene profiles alone. By improving estimation of cross-tissue relationships, SpatialCCA provides a principled framework for integrating two ST slices and can serve as an effective pre-processing step for downstream analysis. 

Keywords

Spatial Transcriptomics

Canonical Correlation Analysis

Dimension Reduction

Cross-tissue Integration 

Speaker

Jingxian Tang, Boston University School of Public Health

Co-Author(s)

Ching-Ti Liu, Boston University School of Public Health
Lukas Weber, Boston University, Department of Biostatistics

SsJSD: A Fusion of Sparsity and Spatial Information for Hi-C Single-Cell Clustering

Single-cell high-throughput chromatin conformation capture (scHi-C) profiles 3D genome architecture at cellular resolution. While recent frameworks use spatial patterns for dissimilarity measures in clustering, the inherent sparsity and high dimensionality of scHi-C matrices pose challenges. Crucially, existing measures often fail to distinguish biologically meaningful structural zeros (SZs) from technical dropouts.
We introduce ssJSD (spatial and structural-zero-aware Jensen-Shannon Divergence), a framework explicitly accounting for scHi-C sparsity. By integrating band-wise contact profiles with SZ-induced sparsity matrices, ssJSD leverages both spatial patterns and biological absence of contacts. We adopted two integration strategies: early fusion, concatenating information into a single representation, and late fusion, integrating JSD-based dissimilarities via diverse averaging. Through simulations and applications to human cell lines and prefrontal cortex data, we demonstrate that ssJSD improves clustering accuracy and effectively distinguishes cell types. Our findings highlight that integrating SZ patterns is important for accurately quantifying cell-to-cell variability. 

Keywords

Single cell Hi-C

Single-cell clustering

Contact distance profile

Structural zeros

Jensen-Shannon divergence

Data integration 

Speaker

Sang Wan Lee, The Ohio State University

Co-Author

Shili Lin, Ohio State University