Transient Receptor Potential Vanilloid 1 (TRPV1) is a tetrameric cation channel expressed in primary afferent neurons, where it contributes to thermosensation and nociception. TRPV1 is a polymodal signal integrator that responds not only to heat, but also to inflammatory agents that produce pain hypersensitivity, including bioactive lipids such as endocannabinoids or lysophosphatidic acid (LPA). Cryo-EM structures have revealed how exogenous ligands, such as capsaicin or drugs, vanilloid bind to and activate TRPV1, but a detailed molecular understanding regarding the actions of endogenous inflammatory lipids remain scarce. Here, we describe how LPA binds to and activates TRPV1 by visualizing multiple ligand-channel substates. These structural data demonstrate that LPA binds cooperatively to TRPV1 and allosterically induces conformational changes that drive channel opening. We These data provide valuable insight into the function of inflammatory lipids on TRPV1 and provides further mechanistic insight into how endogenous agonists activate this channel.