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+ \section*{Introduction}
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+ Bulk RNA sequencing of blood and tissue remains the most widely used transcriptomic assay in clinical research, including large clinical trials, biobanks and disease cohorts. Each bulk profile, however, is a mixture: changes in cell-type abundance are confounded with changes in cell-state-specific expression, and disease signals carried by minority populations are diluted. Single-cell RNA sequencing resolves these components, but cost and sample logistics still limit it to tens or hundreds of donors per study.
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+ Computational deconvolution has been the principal bridge between the two. Digital cytometry methods such as CIBERSORTx\cite{newman2019cibersortx}, MuSiC\cite{wang2019music}, Bisque\cite{jew2020bisque}, DWLS\cite{tsoucas2019dwls}, Scaden\cite{menden2020scaden} and TAPE\cite{chen2022tape} estimate cell-type proportions from bulk using single-cell-derived signatures, and BayesPrism\cite{chu2022bayesprism}, BLADE\cite{barbosa2021blade} and bMIND\cite{wang2021bmind} additionally infer cell-type-specific expression. Despite of the highly advanced calculation algorithms, the accuracy of each model's output highly depends on data transformation, reference choice and annotation granularity, degrades for closely related and rare cell types, and changes rank when the reference differs from the bulk cohort\cite{avilacobos2020benchmarking,sturm2019evaluation,dietrich2026omnideconv,community2024assessment}. These methods also return proportions or cell-type averages, which cannot represent heterogeneity within a cell type, thus masking many disease-associated states that could be inferred by transcriptomics.
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+ Due to the advance in machine learning and neural networking, generative models now offer a route to single-cell resolution. Bulk2Space\cite{liao2022bulk2space} and BulkTrajBlend\cite{zeng2024omicverse} estimate proportions and then draw cells from a matched reference, and scSemiProfiler\cite{wang2024scsemiprofiler} adapts real single cells from actively selected cohort representatives toward each target's bulk profile. These designs either constrain generated populations to a proportion-weighted sum or anchor every generated sample to a representative donor profiled within the same cohort, and their evaluation has relied largely on pseudobulk aggregated from single-cell data\cite{wang2024scsemiprofiler}. In parallel, diffusion models now generate realistic single-cell transcriptomes\cite{luo2024scdiffusion,scdiffusionx2026}, and sparsity-aware formulations that diffuse expression values and their zero pattern jointly preserve the exact zeros that dominate single-cell data\cite{ostheimer2025sparse}. These models are conditioned on labels, not on a donor's bulk transcriptome.
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+ A largely untested assumption shared by these approaches is that measured bulk RNA-seq and single-cell pseudobulk are interchangeable. The two assays differ in library chemistry, transcript coverage, capture efficiency and gene-length sensitivity, and distribution shifts between pseudobulk used for training and real bulk encountered at inference are currently unaccounted for\cite{delafuente2025sweetwater}. If this gap is large and systematic, a model trained on pseudobulk is evaluated in one domain and deployed in another, and the consequences for biological conclusions drawn from generated or deconvolved cells have not been quantified.
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+ Here we present PBISC-Diffusion, a bulk-conditioned generative model that combines joint value--support diffusion in gene space, adapted from Sparse Data Diffusion\cite{ostheimer2025sparse}, with deterministic sampling\cite{song2021ddim} and pretraining on peripheral blood mononuclear cells from 22 independent bulk to single cell paired datasets.
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