Deep learning in spatial transcriptomics: Learning from the next next-generation sequencing
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Abstract
Spatial transcriptomics (ST) technologies are rapidly becoming the extension of single-cell RNA sequencing (scRNAseq), holding the potential of profiling gene expression at a single-cell resolution while maintaining cellular compositions within a tissue. Having both expression profiles and tissue organization enables researchers to better understand cellular interactions and heterogeneity, providing insight into complex biological processes that would not be possible with traditional sequencing technologies. Data generated by ST technologies are inherently noisy, high-dimensional, sparse, and multi-modal (including histological images, count matrices, etc.), thus requiring specialized computational tools for accurate and robust analysis. However, many ST studies currently utilize traditional scRNAseq tools, which are inadequate for analyzing complex ST datasets. On the other hand, many of the existing ST-specific methods are built upon traditional statistical or machine learning frameworks, which have shown to be sub-optimal in many applications due to the scale, multi-modality, and limitations of spatially resolved data (such as spatial resolution, sensitivity, and gene coverage). Given these intricacies, researchers have developed deep learning (DL)-based models to alleviate ST-specific challenges. These methods include new state-of-the-art models in alignment, spatial reconstruction, and spatial clustering, among others. However, DL models for ST analysis are nascent and remain largely underexplored. In this review, we provide an overview of existing state-of-the-art tools for analyzing spatially resolved transcriptomics while delving deeper into the DL-based approaches. We discuss the new frontiers and the open questions in this field and highlight domains in which we anticipate transformational DL applications.
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Self-citationAutomated identification of the mouse brain’s spatial compartments from in situ sequencing data
Partel, G., Hilscher, M. M., Milli, G., Solorzano, L., Klemm, A. H., Nilsson, M. F., & Wählby, C. (2020) · BMC Biology · doi:10.1186/s12915-020-00874-5 · Cited by 33 · see citing works
DEEPsc: A Deep Learning-Based Map Connecting Single-Cell Transcriptomics and Spatial Imaging Data
Maseda, F., Cang, Z., & Nie, Q. H. (2021) · Frontiers in Genetics · doi:10.3389/fgene.2021.636743 · Cited by 33 · see citing works
Semantic segmentation of mFISH images using convolutional networks
Pardo, E., Morgado, J. M., & Malpica, N. (2018) · Cytometry Part A · doi:10.1002/cyto.a.23375 · Cited by 28 · see citing works
Derakhshani, A., Asadzadeh, Z., Safarpour, H., Leone, P., Shadbad, M. A., Heydari, A. A., Baradaran, B., & Racanelli, V. (2021) · Journal of Personalized Medicine · doi:10.3390/jpm11080721 · Cited by 28 · see citing works
Self-citationHuang, X., Li, X., Qin, P., Zhu, Y., Xu, S., & Chen, J. (2018) · Frontiers in Oncology · doi:10.3389/fonc.2018.00582 · Cited by 26 · see citing works
Principles of Spatial Transcriptomics Analysis: A Practical Walk-Through in Kidney Tissue
Noel, T., Wang, Q. S., Greka, A., & Marshall, J. L. (2022) · Frontiers in Physiology · doi:10.3389/fphys.2021.809346 · Cited by 26 · see citing works
Spatially resolved transcriptomics reveals the architecture of the tumor/microenvironment interfacePreprint
Hunter, M. V., Moncada, R. M., Weiss, J. M., Yanai, I., & White, R. M. (2020) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2020.11.05.368753 · Cited by 24 · see citing works
Aragón, J., Eberly, D. H., & Eberly, S. I. (1992) · Statistics & Probability Letters · doi:10.1016/0167-7152(92)90157-z · Cited by 23 · see citing works
Hilscher, M. M., Gyllborg, D., Yokota, C., & Nilsson, M. F. (2020) · Methods in molecular biology · doi:10.1007/978-1-0716-0623-0_20 · Cited by 22 · see citing works
On the Generalization Benefit of Noise in Stochastic Gradient DescentPreprint
Smith, S. L., Elsen, E., & De, S. (2020) · arXiv (Cornell University) · doi:10.48550/arxiv.2006.15081 · Cited by 22 · see citing works
GLUER: integrative analysis of single-cell omics and imaging data by deep neural networkPreprint
Peng, T., Chen, G. M., & Tan, K. (2021) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.01.25.427845 · Cited by 21 · see citing works
Spatial organization of the somatosensory cortex revealed by cyclic smFISHPreprint
Codeluppi, S., Borm, L. E., Zeisel, A., Manno, G. L., van Lunteren, J. A., Svensson, C. I., & Linnarsson, S. (2018) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/276097 · Cited by 19 · see citing works
Flagged first, then by citations
4 of the 212 references couldn’t be matched to a known paper.
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Deep learning applications in single-cell genomics and transcriptomics data analysis
Erfanian, N., Heydari, A. A., Feriz, A. M., Picazo, P. I., Derakhshani, A., GhasemiGol, M., Farahpour, M., Razavi, S. M., Nasseri, S., Safarpour, H., et al. (2023) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2023.115077 · Cited by 162 · see citing works
Self-citationGe, S., Sun, S., Xu, H., Cheng, Q. S., & Ren, Z. (2025) · Briefings in Bioinformatics · doi:10.1093/bib/bbaf136 · Cited by 89 · see citing works
A guidebook of spatial transcriptomic technologies, data resources and analysis approaches
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Microsystem Advances through Integration with Artificial Intelligence
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Deep Learning in Diverse Intelligent Sensor Based Systems
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Deep learning in spatially resolved transcriptomics: a comprehensive technical view
Zahedi, R., Ghamsari, M. R. E., Argha, A., MacPhillamy, C., Beheshti, A., Alizadehsani, R., Lovell, N. H., Lotfollahi, M., & Alinejad‐Rokny, H. (2024) · Briefings in Bioinformatics · doi:10.1093/bib/bbae082 · Cited by 74 · see citing works
Pielawski, N., Andersson, A., Avenel, C., Beháňová, A., Chelebian, E., Klemm, A. H., Nysjö, F., Solorzano, L., & Wählby, C. (2023) · Heliyon · doi:10.1016/j.heliyon.2023.e15306 · Cited by 52 · see citing works
A review on deep learning applications in highly multiplexed tissue imaging data analysis
Zidane, M., Makky, A., Bruhns, M., Rochwarger, A., Babaei, S., Claassen, M., & Schürch, C. M. (2023) · Frontiers in Bioinformatics · doi:10.3389/fbinf.2023.1159381 · Cited by 30 · see citing works
From tissue architecture to clinical insights: Spatial transcriptomics in solid tumor studies
Sharma, A., Raut, S. S., Shukla, A., Gupta, S., Mishra, A., & Singh, A. (2025) · Seminars in Oncology · doi:10.1016/j.seminoncol.2025.152389 · Cited by 29 · see citing works
Role of next‐generation sequencing in revolutionizing healthcare for cancer management
Sahoo, O. S., Aidasani, H., Nayek, A., Tripathi, S., Talukdar, J., Gul, A., Kumar, D., Dhar, R., & Karmakar, S. (2024) · MedComm – Future Medicine · doi:10.1002/mef2.70001 · Cited by 25 · see citing works
Spatial transcriptomic analysis of the mouse brain following chronic social defeat stress
Wang, T., Song, Z., Zhao, X., Wu, Y., Wu, L., Haghparast, A., & Wu, H. (2023) · Exploration · doi:10.1002/exp.20220133 · Cited by 20 · see citing works
Hong, K., Radian, Y., Manda, T., Xu, H., & Luo, Y. (2023) · Plants · doi:10.3390/plants12234006 · Cited by 16 · see citing works
Pielawski, N., Andersson, A., Avenel, C., Beháňová, A., Chelebian, E., Klemm, A. H., Nysjö, F., Solorzano, L., & Wählby, C. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2022.01.28.478131 · Cited by 13 · see citing works
Zhang, M., Zhang, W., & Ma, X. (2024) · Briefings in Bioinformatics · doi:10.1093/bib/bbae437 · Cited by 11 · see citing works
Wu, D., Gaskins, J., Sekula, M. N., & Datta, S. (2023) · Genes · doi:10.3390/genes14071368 · Cited by 8 · see citing works
Miao, Z., Tian, T., Chen, W., Wang, Q., Ma, L., Zhang, D., Xie, M., Yu, Z., Guo, X., Bai, G., et al. (2024) · The Innovation Life · doi:10.59717/j.xinn-life.2024.100097 · Cited by 9 · see citing works
Liu, X., & Ren, X. (2024) · Genomics Proteomics & Bioinformatics · doi:10.1093/gpbjnl/qzae057 · Cited by 7 · see citing works
Spatial transcriptomics meets diabetic kidney disease: Illuminating the path to precision medicine
Liu, D., Hu, H., Li, F.-F., Hu, Q., Liu, M., Hao, Y., & Li, B. (2025) · World Journal of Diabetes · doi:10.4239/wjd.v16.i9.107663 · Cited by 7 · see citing works
Cui, Y., Wei, L., Wang, R., Ye, X., & Sakurai, T. (2024) · Current Bioinformatics · doi:10.2174/0115748936278884240102094058 · Cited by 6 · see citing works
Flexible analysis of spatial transcriptomics data (FAST): a deconvolution approach
Zhang, M., Liu, Y., Parker, J., An, L., & Sun, X. (2025) · BMC Bioinformatics · doi:10.1186/s12859-025-06054-y · Cited by 7 · see citing works
Leng, J., Zhang, Y., Liu, X., Cui, Y., Zhao, J., & Ge, Y. (2023) · IEEE Transactions on Circuits and Systems for Video Technology · doi:10.1109/tcsvt.2023.3301677 · Cited by 6 · see citing works
Liu, T., Huang, T., Ding, T., Wu, H., Humphrey, P., Perincheri, S., Schalper, K., Ying, R., Xu, H., Zou, J., et al. (2025) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2025.01.13.632818 · Cited by 6 · see citing works
Sippl, C., Alicia, S., Teping, F., Vanessa, T., Ketter, R., Jin, K. Y., Linsler, S., & Oertel, J. (2023) · Acta Neurochirurgica · doi:10.1007/s00701-023-05594-7 · Cited by 4 · see citing works
Shah, S. N. A., Aalam, J., & Parveen, R. (2025) · Archives of Computational Methods in Engineering · doi:10.1007/s11831-025-10357-x · Cited by 3 · see citing works
Harnessing spatial transcriptomics to understand host-parasite interactions in plants and animals
Pijnacker, A., Bruggeman, C. W., Korswagen, H. C., Smant, G., & Lozano‐Torres, J. L. (2025) · Biotechnology Advances · doi:10.1016/j.biotechadv.2025.108760 · Cited by 3 · see citing works
Innovation in next-generation sequencing in non-small cell lung cancer diagnostics
Scimone, C., Palumbo, L., Borea, R., Sarracino, C., Tomaiuolo, I., Giovanni, D. D., Alfano, S., Nacchio, M., Russo, G., Russo, A., et al. (2025) · Expert Review of Anticancer Therapy · doi:10.1080/14737140.2025.2549538 · Cited by 2 · see citing works
Lakra, A., Singh, N., Vashishth, S., Ambasta, R. K., & Kumar, P. (2026) · Ageing Research Reviews · doi:10.1016/j.arr.2026.103189 · Cited by 3 · see citing works
N-ACT: An Interpretable Deep Learning Model for Automatic Cell Type and Salient Gene IdentificationPreprint
Heydari, A. A., Davalos, O. A., Hoyer, K. K., & Sindi, S. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2022.05.12.491682 · Cited by 2 · see citing works
Self-citationBoosting Single-Cell RNA Sequencing Analysis with Simple Neural AttentionPreprint
Davalos, O. A., Heydari, A. A., Fertig, E. J., Sindi, S., & Hoyer, K. K. (2023) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2023.05.29.542760 · Cited by 2 · see citing works
Self-citationArtificial intelligence and deep learning in single-cell omics data analysis: A case study
Anjum, Z., Khalid, W., Takkar, G., Chhetri, P., & Raza, K. (2024) · Elsevier eBooks · doi:10.1016/b978-0-443-27523-4.00007-x · Cited by 1 · see citing works
TissuStamp: a novel high-throughput tissue transfer workflow for FFPE spatial biology assays
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Advances in generative biology and bio-inspired artificial intelligence
Wen-Ye, Z. E. N. G., ZHENG, S.-Y., Xiao-Qin, Z. H. A. O., Zhu, W. A. N. G., ZHANG, Y.-J., & Zheng-Yuan, M. A. (2026) · Shengming kexue · doi:10.3724/cbls.2026038 · Cited by 1 · see citing works
The Use of Deep Learning in RNA Therapeutic Development
Subramanian, D. A., Yao, S. L., Chan, A., Witten, J. J., Reker, D., Anderson, D. G., Langer, R., & Traverso, G. (2026) · ACS Nano · doi:10.1021/acsnano.6c02090 · Cited by 1 · see citing works
Deep learning insights into transcriptomics and gene expression patterns analysis
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Bioinformatics and Computational Biology: Tools for the Genomic Era
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Spatial Multi-Omics in Drug Discovery
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Artificial intelligence in spatial transcriptomics: a review of methods, datasets, and directions
Sharma, N., Şen, R., & Erdem, C. (2026) · Briefings in Bioinformatics · doi:10.1093/bib/bbag569
- Authors
- Heydari, A. A., & Sindi, S. S.
- Affiliations
- University of California, Merced (US)
- Venue
- Biophysics Reviews
- Publisher
- AIP Publishing
- Published
- 2023-02-07
- Volume
- 4
- Issue
- 1
- Pages
- 011306-011306
- ISSN
- 2688-4089 (ISSN-L)
- PMC
- PMC10903438
- OpenAlex
- W4319347394
- Cited by
- 44
- References
- 212
- Citations by year
- 2026: 6 · 2025: 13 · 2024: 10 · 2023: 12 · 2022: 3
- FWCI
- 1.08
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- cc-by-nd
- Locations
- Biophysics Reviews (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nd)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (R15HL146779)
- National Science Foundation (DMS-1840265)
- National Human Genome Research Institute (F31HG012718)
- National Institute of General Medical Sciences (R01GM126548)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
- Keywords
- spatial transcriptomics, deep learning, spatial reconstruction, spatial clustering, spatial alignment, multimodal data, tissue organization, gene expression profiling, single-cell RNA sequencing, histological images
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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
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