Work Packages

An integrated framework combining multi-omics, functional neurobiology, and genome editing to dissect metabolic control of puberty.

Overview

The scientific strategy of DOPA-Kiss is articulated into three complementary and interconnected Work Packages (WP1–WP3). Each WP addresses a specific dimension of the project’s objectives, while maintaining strong integration to ensure coherent progress from molecular characterisation to functional validation and causal testing.

This structure enables a logical progression: from comprehensive identification of relevant molecular pathways (WP1), to mechanistic analysis (WP2), and finally to direct experimental demonstration of causality (WP3).

WP1

Multi-omics characterisation of hypothalamic kisspeptin and dopaminergic neurons

Aim:

To define the molecular, transcriptional, and metabolic profiles of hypothalamic kisspeptin and dopaminergic neurons under normal conditions and in models of early-onset obesity.

Key approaches:

✅ Comprehensive metabolomic and lipidomic profiling of the hypothalamus.

✅ Isolation of kisspeptin and dopaminergic neurons for pooled and single-cell transcriptomics and epigenomics.

✅ Spatial multi-omics to map metabolic alterations in situ.

Expected outcome:

A detailed molecular atlas identifying deregulated pathways and potential targets for functional testing in WP2 and WP3.

WP2

Mechanistic analysis of metabolic modulation

Aim:

To investigate how nutrient- and lipid-sensing pathways, stress responses, and autophagic mechanisms influence the function of kisspeptin and dopaminergic neurons.

Key approaches:

✅ Quantitative analysis of gene and protein expression in whole tissue and sorted neurons.

✅ Central pharmacological interventions targeting metabolic sensors.

✅ Functional manipulation using chemogenetics and optogenetics in vivo.

Expected outcome:

Identification of specific molecular mechanisms by which metabolic cues influence pubertal timing, including sex-specific effects.

WP3

Causal validation through targeted genome editing

Aim:

To establish direct causal relationships between identified molecular pathways and the regulation of puberty and metabolism.

Key approaches:

✅ CRISPR/Cas9-mediated editing in kisspeptin neurons and associated pathways.

✅ Stereotaxic delivery of viral vectors for spatially targeted gene manipulation.

✅ Phenotypic analysis of pubertal timing and metabolic parameters.

Expected outcome:

Definitive experimental evidence linking specific molecular targets to the central control of puberty.