Figure 4: A circuit-first evidence-gated agentic workflow for cross-scale target discovery.A user initiates a campaign with a central nervous system disease and therapeutic modality, after which subsequent inputs may be provided in free form. A coordinating PI agent manages the workflow, applies gates, identifies failures, and re-triggers earlier stages when evidence is insufficient. The circuit agent identifies disease-relevant activity phenotypes, circuits, regions, and cell populations using CircuitATLAS. After a gate requiring sourced and directionally coherent evidence, the molecular agent searches for effectors capable of modulating the selected circuit phenotype while direct disease-to-molecule information is masked to prevent shortcut reasoning. The modality agent evaluates the feasibility of engaging the proposed target, including intervention, vector, promoter, and campaign-level constraints, and the clinical agent assesses existing trials, biomarkers, safety, and translational evidence. An adversarial sceptic independently challenges the reasoning produced at each specialist stage. Three memory layers maintain the read-only semantic knowledge graph, an episodic campaign graph written by the PI agent, and a compacted working transcript. The workflow terminates in ranked target dossiers containing the supporting evidence, assumptions, uncertainties, and proposed route to validation; dossiers that fail predefined evidence or feasibility gates are rejected or returned for further search.To illustrate the hypotheses generated by this workflow, we applied the agent to Fragile X syndrome, Dravet syndrome and epilepsy (Table 4). In Fragile X syndrome, the workflow identified excessive thalamocortical gain and nominated partial suppression of SLC17A6 (VGLUT2) in excitatory thalamic relay neurons as a corrective intervention. Reducing vesicular glutamate loading was predicted to lower quantal transmission at thalamocortical synapses and thereby reduce cortical gain, although the essential role and broad subcortical expression of VGLUT2 make partial and anatomically restricted modulation an important constraint. In Dravet syndrome, the agent proposed partial suppression of the somatodendritic HTR1A autoreceptor in dorsal raphe serotonergic neurons, with the aim of releasing these neurons from autoinhibition and increasing forebrain serotonin tone. This strategy has clinical precedent in the indication, since fenfluramine is approved for seizures associated with Dravet syndrome and acts in part through serotonergic mechanisms, although the workflow also surfaced potentially divergent effects of 5-HT1A signalling across seizure phenotypes. In epilepsy, CircuitATLAS nominated ATP1A3, the neuronal α\\alpha3 Na+/K+-ATPase, as a control point on cortical excitability, with potentiation predicted to increase activity-dependent hyperpolarizing current and preserve inhibitory restraint during sustained firing. We selected ATP1A3 for experimental validation and carried this hypothesis into the in vivo and structure-guided campaign described in the next section. Together, these examples illustrate how CircuitATLAS identifies targets through their capacity to control pathological circuit states rather than through their molecular association with disease. 出典: CircuitATLAS: Agentic reasoning over a systems neuroscience knowledge graph for target discovery in circuitopathies（arXiv）Figure 5: An in vivo lab-in-the-loop, from a graph-derived control point to a chemistry campaign.(A) Campaigns enter at one of four points and are planned by an experiment-design agent that addresses the laboratory through a Model Context Protocol server, exposing 25 tools and 20 resources over a structured record of 81 versioned procedures, operator competence, equipment state, colony and consumable stock, reserved capacity and cost per subject. Designs pass deterministic gates before human approval; an assay the laboratory does not hold enters as a capability request. (B) Pump turnover exports net positive charge and depends on intracellular Na+, so the hyperpolarizing current it generates is recruited in proportion to the Na+ that firing accumulates (inset). (C) Mean cortical LFP power spectra for AAV9.mDlx.ATP1A3 and the reference vector at baseline and under focal 4-AP, canonical bands shaded. Baseline power differs between arms, so the primary contrast is within-animal. (D) Absolute band power, one point per animal, with medians (reference baseline n=8n=8, ATP1A3 baseline n=6n=6, reference 4-AP n=5n=5, ATP1A3 4-AP n=7n=7); qq values are Benjamini–Hochberg corrected across bands for the within-animal fold-change contrast. (E) Structure-guided campaign on the validated target, with one representative compound and predicted binding score per site, sites ordered by the prior probability that occupancy raises turnover. Docking predicts binding, not polarity. (F) The polarity screen: 48 wells across the four site classes, thallium flux on human α\\alpha3 with α\\alpha1 and α\\alpha2 in parallel on an ouabain-resistant background, at sub-maximal Na+ load.We used ATP1A3 to illustrate how a target hypothesis can be advanced through this process. CircuitATLAS nominated the neuronal α\\alpha3 Na+/K+-ATPase as a control point on cortical excitability. Pump turnover exports net positive charge and is recruited by the intracellular Na+ load that firing generates, producing an activity-dependent hyperpolarizing current (Figure 5B)36. ATP1A3 is restricted to neurons and strongly expressed in GABAergic populations37, while loss of function is epileptogenic38 and pharmacological pump inhibition is convulsant39, together suggesting potentiation as the therapeutic direction. The unresolved question was whether increasing ATP1A3 in inhibitory interneurons would strengthen inhibition during sustained firing or instead suppress their output. 出典: CircuitATLAS: Agentic reasoning over a systems neuroscience knowledge graph for target discovery in circuitopathies（arXiv）
