OBJECTIVES/GOALS: We aim to discover safer and more effective therapeutics for CNS disorders. Current therapeutic development is hindered by dosing out drugs for safe consumption. By identifying proteins with narrow functional roles in the brain (i.e., behavioral control), we can develop drugs targeting these proteins for improved treatment safety and efficacy. METHODS/STUDY POPULATION: We focused on an evolutionarily new, non-neuronal, non-synaptic glutamate signaling mechanism, system xc- (Sxc). Sxc activity was eliminated by mutating the gene Slc7a11 through pronuclear injection of zinc-finger nucleases into Sprague Dawley rat embryos to create a line of rats lacking Sxc (MSxc). To confirm Sxc mutation, we verified that tissue from MSxc rats had a complete lack of xCT, which is the regulatory subunit of Sxc that is encoded by Slc7a11. We also verified that astrocyte cultures generated from MSxc tissue lacked cystine-evoked glutamate release. Next, we measured development (body weight), CNS regulation of metabolism, and other indicators of generalized, non-specific brain function as well as behaviors that are reliant on behavioral control, such as impulse control and response inhibition. RESULTS/ANTICIPATED RESULTS: Eliminating Sxc was not lethal and did not impair development or produce widespread changes in brain function as is commonly observed when deleting other glutamate mechanisms. MSxc rats did not differ from wildtype in growth rate, central regulation of metabolism as reflected by absolute or diurnal changes in core body temperature, locomotor activity in a familiar or novel environment, or simple forms of cognition such as novel object recognition, or operant responding (food and cocaine-reinforced). In contrast, behaviors that rely on behavioral control were impaired. MSxc rats displayed deficits in impulse control and behavioral flexibility. We hypothesize that MSxc rats will also show deficits in response inhibition using the stop signal reaction time task, a common metric used in clinical populations. DISCUSSION/SIGNIFICANCE: Eliminating Sxc activity in rats produced deficits in behaviors reliant on impulse control, without impacting development or simple brain function. These results show the potential of targeting Sxc to restore behavioral control without generating therapeutically limiting adverse effects resulting from non-specific changes in brain function.

OBJECTIVES/GOALS: We aim to determine whether non-neuronal, non-synaptic glutamate signaling mechanisms can be targeted to produce highly specific, narrow changes in brain function that would benefit CNS disorders. To do this, we investigated cognitive changes produced through manipulating the activity of the astrocytic glutamate release mechanism system xc-. METHODS/STUDY POPULATION: System xc- (Sxc) activity was eliminated by mutating the gene Slc7a11 through pronuclear injection of zinc-finger nucleases into Sprague Dawley rat embryos to create a line of rats lacking Sxc (MSxc rats). To confirm a lack of Sxc activity, we verified that tissue from MSxc rats had a complete lack of xCT, which is the regulatory subunit of Sxc that is encoded by Slc7a11. We also verified that astrocyte cultures generated from MSxc tissue lacked cystine-evoked glutamate release. Next, we measured development (body weight), CNS regulation of metabolism, and other indicators of generalized, non-specific brain function as well as behaviors that are reliant on executive function, such as cognitive flexibility, impulse control, decision-making, and response inhibition. RESULTS/ANTICIPATED RESULTS: Eliminating Sxc was not lethal and did not impair development or produce widespread changes in brain function as is commonly observed when deleting other glutamate mechanisms. MSxc rats did not differ from wildtype in growth rate, central regulation of metabolism as reflected by absolute or diurnal changes in core body temperature, locomotor activity in a familiar or novel environment, or simple forms of cognition such as novel object recognition, or operant responding (food and cocaine-reinforced). In contrast, behaviors that rely on executive function were impaired. MSxc rats displayed deficits in cocaine reinstatement and attentional set-shifting. We anticipate MSxc rats to also show impairments in decision-making in the rat gambling task and response inhibition in the stop-signal reaction time task. DISCUSSION/SIGNIFICANCE: Eliminating Sxc activity in rats produced deficits in behaviors reliant on executive function without impacting development or simple brain function. These results highlight the potential of targeting Sxc to enhance cognition without generating therapeutically limiting adverse effects resulting from non-specific changes in brain function.

Recent publications concerned with attitudes to the human body in the religion of Mani have revealed a complex spectrum of ideas. A reading of the “Manichaean body” informed by a gnostic polarity of flesh versus spirit has been largely rejected, and a more complex, ambivalent portrayal of the body, shaped by specific cosmological and theological readings of its origin and purpose, has come to light. New interpretive tools and approaches have changed perceptions of classical texts and revealed how the “subjugated, perfected [Manichaean body was] put into use in the process of salvation.” For example, rereading chapter 70 of the Coptic work the Kephalaia of the Teacher, we encounter a complex lesson that betrays the Manichaeans’ understanding of the dual heritage of the human body. Here the Mani of the Kephalaia instructs his disciples about the correspondences that exist between the fleshly body and the universe and formulates them in a manner that suggests a simultaneous patterning of the two forms: “Mani says to his disciples: ‘This whole universe, above and below, reflects the pattern of the human body; as the formation of this body of flesh accords to the pattern of the universe’” (70.169.28–170.1). The organs and limbs of the body resemble specific astral structures and elements in the universe, and both body and universe are afflicted by a range of competing powers. Chapter 70 offers a melothesiac reading of these archontic powers as zodiacal signs fused with the organs, bones, and sinews of the body (cf. chapter 69). As archons they exercise a malevolent influence over the flesh. However, they are also constantly in conflict with each other, and the cause of bodily sickness lies in their “creeping, and moving within the body. . . [where] they shall beset and destroy one another. . . they shall erupt from the body of the person who will die; and make putrid boils and sores and burning wounds in the body” (70.175.12–14, 16–18). Leaving such colorful descriptions of lesions aside, chapter 70 also indicates that human beings, specifically the Manichaean elect, possess enormous potential as the ones who are able to facilitate the release of the “light” by subduing the activities of the “five camps” (i.e., the face, heart, genitalia, stomach, and ground).