AnnoTuneAgent

Documentation: Read the Docs.

Cluster-level cell-type annotation for single-cell RNA-seq, with two fine-tuned language models that run locally.

Each cluster gets a marker score, a name-model result, and an ML-model result. A rule-based judgment reads all three and writes the title, so every call stays auditable.

What is fine-tuned

One Qwen3.5-4B base (Q6_K GGUF) is shared by two independent LoRA adapters. The adapters are not merged into the base, so they load together on a single local llama-server and can be replaced without touching the base.

Both models are trained to copy a name from the candidates they are shown, not to free-write one. A name outside the candidate list cannot become a title, and a name with no marker that passes expression and specificity validation is flagged (no_marker_passed) instead of being trusted silently.

The weights are on the Hugging Face Hub: lieberlee/annotune-qwen3.5-4b-gguf (preview). Inference is local: no cluster data leaves the machine.

Pipeline

raw counts ─ prepare ─▶ QC, doublets, normalization, batch correction, Leiden
           ─ run ─────▶ per cluster:
                          1. marker score        (built-in human / mouse dictionary)
                          2. name model          (LoRA, copies from dictionary_hits)
                          3. ML model            (LoRA, copies from measured candidates)
                          ▼
                        judgment ─▶ title + status + reason + hint
                          ▼
                        validated marker panels, dotplot, report, annotated h5ad

• Judgment, not voting. Agreement among the three results gives a clear title. Disagreement, mixed programs, and unvalidated names are kept and flagged with a reason (mixed_programs, near_clear_cluster, subcluster_labels, no_marker_passed) so you can review them.
• Marker evidence is checked, not quoted. Panels are selected from cited dictionary markers and validated against measured expression and specificity in your data.
• Every title is traceable. The report records each model's answer, the panel, and the reason a cluster was or was not settled.

The models run when models is passed (or the CLI adapter flags are given). Without them, run uses the marker score alone.

Install

Python 3.10–3.13.

pip install annotuneagent

Local GGUF models need pip install 'annotuneagent[models]'. Optional methods: AmbiDose (denoise=True, install), scVI and SOLO (annotuneagent[scvi]), scFair (annotuneagent[scfair]), Harmony, and Scanorama. If you select one that is missing, prepare prints the install command and leaves the matrix unchanged. See Installation.

From a checkout:

git clone https://github.com/leelieber2025/AnnoTuneAgent.git
cd AnnoTuneAgent
pip install -e ".[dev]"

First run

prepare expects raw integer counts. run expects finite, nonnegative, library-size-normalized natural log1p expression with Scanpy's adata.uns["log1p"], which prepare writes.

import annotuneagent as ata

adata = ata.read_10x("outs", ["sample_a", "sample_b"])
adata = ata.prepare(adata, species="human", batch_key="sample")
report = ata.run(adata, "leiden", species="human", subcluster=False)

read_10x takes one filtered matrix per sample folder (*.h5, or the mtx directory when there is no h5). Scrublet and SOLO run per sample on those filtered counts. doublet_chip="10k" (default) uses 0.8% multiplets per 1,000 recovered cells, and "20k" uses 0.4%.

To score a caller-supplied list instead of the built-in dictionary:

records = ata.annotate(
    adata,
    cluster_key="leiden",
    hypotheses=[
        ata.Hypothesis(
            "CD4-positive, alpha-beta T cell",
            positive=("CD3E", "CD4"),
            negative=("CD8A",),
        ),
        ata.Hypothesis(
            "CD8-positive, alpha-beta T cell",
            positive=("CD3E", "CD8A"),
            negative=("CD4",),
        ),
    ],
)

A label needs positive marker enrichment. A negative marker counts against a hypothesis only when the cluster is enriched for it relative to the other cells. Scores are operational, not probabilities.

The built-in table is markers.tsv: human and mouse, each with a tissue-agnostic core and organ pages. Tissue rows stay hidden until tissue matches. Symbols are not translated across species. evaluate compares records with a per-cell author label: the reference for a cluster is the majority label, a tied majority is left unmatched, and a parent name counts as a coarse match.

annotuneagent \
  --input clustered.h5ad \
  --cluster-key leiden \
  --species human \
  --output result.json

--name-adapter, --ml-adapter, and --base run the name and ML models on one local server and require --output-h5ad. The weights are on the Hugging Face Hub: lieberlee/annotune-qwen3.5-4b-gguf. The tokenizer is installed with the package. See docs/tutorials/load_model.ipynb.

Status

0.1.1 (Alpha). Import as import annotuneagent as ata.

Citation

For the software record, cite the Zenodo DOI. Pin the package version used in the analysis, for example annotuneagent==0.1.1.

License

Software: Apache License 2.0.

Author

Zhao Li (李钊) Email: leelieber@gmail.com