Some 40 million patients are given general anesthesia each year in the USA using agents that haven't changed for decades. General anesthetics are used as hypnotics during procedures, sedatives in the ICU, and as anticonvulsants. These drugs have low safety margins and are particularly harmful to newborns, the severely ill, and the elderly. The molecular mechanisms of general anesthesia have proved difficult to reveal, hampering the design of improved agents. This interdisciplinary PPG brings together a group of experienced scientists to focus on mechanisms underlying the ability of general anesthetics to enhance the activity of the inhibitory GABAA receptors (GABAAR) and glycine receptors (GlyR), major targets of anesthetics. The overall hypothesis is that the various actions of general anesthetics are mediated by a number of distinct binding sites on these receptors, that their location and affinity varies with the receptor's subunit composition and conformational state. The overall aims of the PPG are to: (i) locate the anesthetic binding sites on heteromeric GABAARs and GlyRs using anesthetic photolabels, (ii) define anesthetic structure-activity relationships at distinct sites, and (iii) define how their occupancy allostericlly modulates receptor function using electrophysiology and targeted mutations. Project 1 (PI: Cohen) will locate the unknown binding sites for steroids and propofol anesthetics and a convulsant barbiturate on synaptic alpha-1-beta-3 gamma-2 and extrasynaptic alpha-4-beta-3-delta GABAARs and on alpha-1-beta GlyRs. Project 2 (PI: Miller) focuses on exploring the molecular pharmacology of the different anesthetic binding sites that the PPG has discovered on alpha-1-beta-3-gamma-2 and alpha-4-beta-3-delta GABAARs, with the objective of understanding how to improve the specificity of anesthetic action on synaptic and extrasynaptic receptors. In addition, new binding sites that have high affinity for the activated states will be sought. Project 3 (PI: Forman) will use mutations and electrophysiology to define the functional roles, including drug specificity, of distinct general anesthetic binding sites, particularly the iter-subunit transmembrane sites on alpha-1-beta-3-gamma-2 and alpha-4-beta-3-delta GABAARs and alpha-1-beta GlyRs. Photolabeling results and homology models will guide mutagenesis, while allosteric co-agonist modeling will be used as a quantitative analytical framework for electrophysiological data. The Projects are supported by four Core facilities. An Administrative Core (Core A;PI: Miller) coordinates the activities of the components of the PPG, also arranging external oversight through a distinguished Advisory Board. A Synthetic Chemistry Core (Core B;PI: Bruzik) develops and supplies novel general anesthetics for photolabeling and structure activity relationships. A Protein Chemistry Core (Core C;PI: Cohen) locates anesthetic sites of photoincorporation and develops homology models of receptors to guide research. A Protein Production Core (Core D;co-PIs: Miller &Forman) supplies nanomoles of heterologously expressed, purified and functionally reconstituted human neuronal receptors for photolabeling and other studies.

Public Health Relevance

General anesthetics are essential to modern medicine, yet they have low safety margins especially in sick or elderly patients, requiring delivery by highly trained clinicians. General anesthetics interact with many receptors in the brain to cause both anesthesia and side effects. This Program Project aims to provide new knowledge so that new anesthetics can target just the anesthetic receptors and not those responsible for side effects.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
2P01GM058448-16
Application #
8742127
Study Section
Special Emphasis Panel (ZGM1)
Program Officer
Cole, Alison E
Project Start
1998-12-01
Project End
2019-08-31
Budget Start
2014-09-01
Budget End
2015-08-31
Support Year
16
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Feng, Hua-Jun; Forman, Stuart A (2018) Comparison of ??? and ??? GABAA receptors: Allosteric modulation and identification of subunit arrangement by site-selective general anesthetics. Pharmacol Res 133:289-300
McGrath, Megan; Ma, Celena; Raines, Douglas E (2018) Dimethoxy-etomidate: A Nonhypnotic Etomidate Analog that Potently Inhibits Steroidogenesis. J Pharmacol Exp Ther 364:229-237
Zhou, Xiaojuan; Desai, Rooma; Zhang, Yinghui et al. (2018) High-level production and purification in a functional state of an extrasynaptic gamma-aminobutyric acid type A receptor containing ?4?3? subunits. PLoS One 13:e0191583
Ziemba, Alexis M; Szabo, Andrea; Pierce, David W et al. (2018) Alphaxalone Binds in Inner Transmembrane ?+-?- Interfaces of ?1?3?2 ?-Aminobutyric Acid Type A Receptors. Anesthesiology 128:338-351
Forman, Stuart A (2018) Combining Mutations and Electrophysiology to Map Anesthetic Sites on Ligand-Gated Ion Channels. Methods Enzymol 602:369-389
Woll, Kellie A; Zhou, Xiaojuan; Bhanu, Natarajan V et al. (2018) Identification of binding sites contributing to volatile anesthetic effects on GABA type A receptors. FASEB J 32:4172-4189
McGrath, Megan; Yu, Zhiyi; Jayakar, Selwyn S et al. (2018) Etomidate and Etomidate Analog Binding and Positive Modulation of ?-Aminobutyric Acid Type A Receptors: Evidence for a State-dependent Cutoff Effect. Anesthesiology 129:959-969
Ma, Celena; Pejo, Ervin; McGrath, Megan et al. (2017) Competitive Antagonism of Anesthetic Action at the ?-Aminobutyric Acid Type A Receptor by a Novel Etomidate Analog with Low Intrinsic Efficacy. Anesthesiology 127:824-837
Jounaidi, Youssef; Cotten, Joseph F; Miller, Keith W et al. (2017) Tethering IL2 to Its Receptor IL2R? Enhances Antitumor Activity and Expansion of Natural Killer NK92 Cells. Cancer Res 77:5938-5951
Yu, Zhiyi; Cohen, Jonathan B (2017) Enantiomeric barbiturates bind distinct inter- and intrasubunit binding sites in a nicotinic acetylcholine receptor (nAChR). J Biol Chem 292:17258-17271

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