Identification and functional characterization of ion channel-multiprotein complexes involved in nociception
Final Report Abstract
The aim of our research is to enhance our knowledge about the molecular signature of somatosensation in general and specifically of distinct pain states in vertebrates. Pain is a major symptom of many medical conditions and the worldwide number one reason for people to seek medical assistance. It affects the quality of life of patients, and poses a heavy financial burden on society with high costs of treatment and lost productivity. Furthermore, treatment of chronic pain conditions presents a big challenge as pain therapeutics often lack efficacy and exhibit only minimal safety profiles. The latter can be largely attributed to the fact, that current therapies target molecules with key physiological functions throughout the body. In light of these difficulties, the identification of proteins specifically involved in chronic pain states is of paramount importance for designing selective interventions. To this end we use two complementary strategies: On one hand a “candidate”-approach focused on elucidating the interactome of three ion channels crucially involved in nociception (TRPA1 and TRPV1) and mechanosensation (Piezo2) in vertebrates (please see my previous report). On the other hand we successfully extended our research beyond ion channels and opened a new line of investigation with the aim to decipher system-wide changes in protein networks of DRG during different pathological pain conditions. Both these efforts have already yielded exciting, unexpected and novel insights into the molecular basis of somatosensation and specific pain states: A) Our work on TRPA1 channels revealed that they are tightly regulated by its binding partner Anxa2. Using in vitro and in vivo studies we could show that Anxa2 limits TRPA1 membrane trafficking with concomitant effects on TRPA1 function and TRPA1-dependent pain behaviors in mice. Importantly, this regulation is specific for TRPA1 channels while TRPV1 channels, which are co-expressed with TRPA1, are not affected. Hence, we uncovered a mechanism to specifically regulate TRPA1-mediated nociceptive signaling. For this work my PhD student Luca Avenali got awarded with the “Förderpreis Schmerzforschung (Dt. Schmerzgesellschaft e.V.); category: basic research” in 2015. Further investigations of TRPA1 and TRPV1 channels in my laboratory defined a set of novel interaction partners in dorsal root ganglia. Intriguingly, we demonstrated the validity of our initial hypothesis, i.e. that a certain set of TRPA1- or TRPV1-associated proteins are differentially regulated during inflammatory pain and that pain-specific interactions can be identified. The validation of these findings for TRPV1 has been subject of research during the prolongation year of Emmy Noether funding. These experiments were successful and led to the publication of our work in PAIN. In addition, we included a novel subproject into our work schedule and successfully identified Piezo2-associated protein complexes in dorsal root ganglia. These results open the possibility to gain insights into vertebrate mechanotransduction on the molecular level – a long-sought goal of the somatosensory research community. B) In parallel and complementary to the above mentioned projects, we effectively established an integrated workflow, which provided us with an unbiased, quantitative and global view of the protein dynamics underlying chronic pain states of different etiology, inflammatory and neuropathic pain. We determined differential and pain-associated regulation of both, proteins previously linked to pain syndromes in humans and rodents as well as dozens of proteins that have not been described in the context of pain before. These candidate proteins will serve as a stepping-stone for further studies (within the research community and in my laboratory) to elucidate yet unknown signaling pathways relevant and specific for chronic pain (work in progress). On the basis of these results I got awarded with the “Max von Frey-Preis” for pain research (Dt. Schmerzgesellschaft e.V.). Furthermore, we assembled our proteome data of the pain neuraxis in a freely accessible online database serving the scientific community: http://painproteome.em.mpg.de/
Publications
- (2019) Vti1b promotes TRPV1 sensitization during inflammatory pain. Pain 160 (2) 508–527
Sondermann, Julia R.; Barry, Allison M.; Jahn, Olaf; Michel, Niklas; Abdelaziz, Reham; Kügler, Sebastian; Gomez-Varela, David; Schmidt, Manuela
(See online at https://doi.org/10.1097/j.pain.0000000000001418) - Avenali L, Narayanan P, Rouwette T, Cervellini I, Sereda M, Gomez-Varela D, Schmidt M. (2014). Annexin A2 Regulates TRPA1-Dependent Nociception. J Neurosci. 34(44):14506-16
Avenali L, Narayanan P, Rouwette T, Cervellini I, Sereda M, Gomez-Varela D, Schmidt M
(See online at https://doi.org/10.1523/JNEUROSCI.1801-14) - (2016). Native Piezo2 Interactomics Identifies Pericentrin as a Novel Regulator of Piezo2 in Somatosensory Neurons. J Proteome Res, 5;15(8):2676-87
Narayanan P, Sondermann J, Rouwette T, Karaca S, Urlaub H, Mitkovski M, Gomez-Varela D, Schmidt M
(See online at https://doi.org/10.1021/acs.jproteome.6b00235) - (2016). Nocistatin sensitizes TRPA1 channels in peripheral sensory neurons. Channels (Austin). 2016 Jun 30:1-9
Avenali L, Abate Fulas O, Sondermann J, Narayanan P, Gomez-Varela D, Schmidt M
(See online at https://doi.org/10.1080/19336950.2016.1207025) - (2016). Standardized profiling of the membrane-enriched proteome of mouse dorsal root ganglia provides novel insights into chronic pain. Molecular & Cellular Proteomics
Rouwette T, Sondermann J, Avenali L, Gomez-Varela D, Schmidt M
(See online at https://doi.org/10.1074/mcp.M116.058966) - (2017). Anti-nociceptive action of peripheral mu-opioid receptors by G-beta-gamma protein-mediated inhibition of TRPM3 channels. eLife
Sandeep Dembla, Marc Behrendt, Florian Mohr, Christian Goecke, Julia Sondermann, Franziska M Schneider, Marlene Schmidt, Julia Stab, Raissa Enzeroth, ... Dominik Oliver, Eleonora Zakharian, Manuela Schmidt, Johannes Oberwinkler
(See online at https://doi.org/10.7554/eLife.26280.001) - (2017). New targeted approaches for the quantification of data-independent acquisition mass spectrometry. Proteomics
Bruderer R, Sondermann J, Tsou CC, Barrantes-Freer A, Stadelmann C, Nesvizhskii AI, Schmidt M, Reiter L, Gomez-Varela D
(See online at https://doi.org/10.1002/pmic.201700021)
