Cellular targets for treating reduced cognitive flexibility in obsessive compulsive disorder and uses thereof

Abstract

The application discloses a cell target for treating reduced cognitive flexibility of obsessive-compulsive disorder and belongs to the technical field of biotechnology, and comprises the following steps: locating a functionally abnormal time window and an event type; verifying whether correction or simulation of the functionally abnormal time window can improve or induce cognitive flexibility disorder, and establishing a cause-effect relationship; verifying that functionally abnormal intermediate neurons in specific subtypes of specific brain regions are the root cause of the cognitive flexibility disorder, and determining the cell target for treating reduced cognitive flexibility of obsessive-compulsive disorder; the application first identifies SST subtype intermediate neurons and PV subtype intermediate neurons in the lateral orbitofrontal cortex as specific cell targets for treating reduced cognitive flexibility of obsessive-compulsive disorder, and the two types of intermediate neurons have different functional roles in cognitive flexibility: functionally abnormal SST subtype intermediate neurons selectively impair the ability of new rule exploration, and functionally abnormal PV subtype intermediate neurons impair the abilities of new rule exploration and maintenance.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder and its application. Background Technology

[0002] Obsessive-compulsive disorder (OCD) is a common chronic neuropsychiatric disorder. Patients typically exhibit intrusive, persistent thoughts or repetitive, stereotyped compulsive behaviors. These behaviors consume a significant amount of time, cause substantial psychological distress, and severely interfere with an individual's daily work, life, and social functioning.

[0003] Current treatments for obsessive-compulsive disorder (OCD) primarily rely on medication and deep brain stimulation (DBS), which present the following problems: 1. Lack of cell specificity in target: It is impossible to distinguish different neuronal subtypes and act on all neurons in the entire brain region, resulting in a lack of precision in treatment and possible unnecessary side effects; 2. Unclear Mechanisms and Unstable Efficacy: Due to the lack of accurate understanding of the specific cell types and neural circuits mediating cognitive flexibility disorders, the mechanisms of action of existing treatments are unclear, and the efficacy varies greatly from person to person and is difficult to predict. 3. Inability to distinguish cognitive sub-components: It is impossible to differentiate and target different cognitive deficits such as "impaired ability to explore new rules" and "impaired ability to maintain new rules," resulting in a lack of functional specificity in treatment; 4. Limited by critical developmental period: Some interventions need to be implemented in the early stages of development, which makes them less applicable to adult patients.

[0004] In view of this, we designed a cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder and its application to address the aforementioned problems. Summary of the Invention

[0005] To address the problems mentioned in the background section, this invention provides a cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder and its application, characterized by a clearly defined cellular target, strong functional specificity, and the ability to be intervened in adulthood.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder (OCD), comprising the following steps: First, using fiber optic imaging technology, the dynamic changes in neuronal activity of SST and PV subtype interneurons in the lateral orbitofrontal cortex of an OCD animal model and a normal control animal during a reverse learning task are recorded to locate the time window and event type of functional abnormality; based on the determined time window of functional abnormality, the activity of specific subtype interneurons is specifically regulated using optogenetics to verify whether correcting or simulating functional abnormality can improve or induce cognitive flexibility impairment, thereby establishing a causal relationship; using conditional gene manipulation technology, the Sapap3 gene is specifically knocked out or restored in specific subtype interneurons in the lateral orbitofrontal cortex of adult animals to verify that functional abnormality of specific subtype interneurons in specific brain regions is the root cause of cognitive flexibility impairment, thus determining the cellular target for treating cognitive flexibility impairment in OCD, wherein the cellular target is the SST and PV subtype interneurons in the lateral orbitofrontal cortex.

[0007] Furthermore, the location of the time window and event type of the functional abnormality includes: for SST subtype interneurons, in the early stage of reversal learning, during the presentation of cues with rewards before reversal but no rewards after reversal and the behavioral selection stage, neuronal activity is abnormally reduced; for PV subtype interneurons, in the early and late stages of reversal learning, during the presentation of cues for incorrect selection events after the presentation of no-reward odors and the result feedback stage, neuronal activity is abnormally reduced.

[0008] Furthermore, the optogenetic technique-specific regulation includes: for SST subtype interneurons, specific regulation is performed on the 0-3 second cue presentation phase after rule reversal where there is a reward before reversal but no reward after reversal; for PV subtype interneurons, specific regulation is performed on the 0-3 second cue presentation phase and the 6-9 second result feedback phase in trials without reward after rule reversal.

[0009] Furthermore, the conditional gene manipulation technology includes: specifically knocking out the Sapap3 gene in interneurons of the SST or PV subtypes of the lateral orbitofrontal cortex of adult mice, and observing whether it induces cognitive flexibility impairment; then specifically re-expressing the Sapap3 gene in interneurons of the SST or PV subtypes of the lateral orbitofrontal cortex of adult mice, and observing whether it salvages cognitive flexibility impairment.

[0010] Application of a cellular target for treating cognitive flexibility impairment in the preparation of drugs or interventions for treating cognitive flexibility disorders.

[0011] Furthermore, the application of the SST subtype interneurons is as follows: to treat cognitive flexibility impairment, which is mainly manifested by impaired ability to explore new rules, with the intervention time window being the early stage of rule reversal.

[0012] Furthermore, the application of the PV subtype interneurons is as follows: to treat cognitive flexibility impairment that simultaneously impairs the ability to explore new rules and the ability to maintain new rules, with the intervention time window covering the early and late stages of rule reversal.

[0013] Furthermore, the restoration of cognitive flexibility through targeted intervention of specific inhibitory interneuron subclasses in the lateral orbitofrontal cortex includes optogenetic regulation and gene therapy.

[0014] Furthermore, the targeted intervention is implemented in adulthood and is not limited to critical developmental periods.

[0015] Compared with the prior art, the beneficial effects of the present invention are: 1. This invention is the first to identify SST subtype interneurons and PV subtype interneurons in the lateral orbitofrontal cortex as specific cellular targets for treating cognitive flexibility impairment in obsessive-compulsive disorder. The two have different functional roles in cognitive flexibility: abnormal function of SST subtype interneurons selectively impairs the ability to explore new rules, while abnormal function of small PV subtype interneurons impairs the ability to explore and maintain new rules.

[0016] 2. The target discovery method established in this invention is not limited by the critical developmental period and is applicable to adult intervention, providing a theoretical basis for the development of precision targeted therapy. Attached Figure Description

[0017] Figure 1 This is a schematic diagram illustrating the dynamic activity of SST subtype interneurons in the lateral orbitofrontal cortex during inversion learning, as recorded by fiber optic calcium imaging according to the present invention. Figure 2 This is a schematic diagram illustrating the dynamic activity of PV subtype interneurons in the lateral orbitofrontal cortex during inversion learning, recorded by fiber optic calcium imaging according to the present invention. Figure 3 This is a schematic diagram illustrating the causal relationship between abnormal interneuron function of the SST subtype in the lateral orbitofrontal cortex and decreased cognitive flexibility in obsessive-compulsive disorder, as verified by optogenetic analysis in this invention. Figure 4 This is a schematic diagram illustrating the causal relationship between abnormal function of PV subtype interneurons in the lateral orbitofrontal cortex and reduced cognitive flexibility in obsessive-compulsive disorder, as verified by optogenetic analysis in this invention. Figure 5 This is a schematic diagram illustrating how the conditional gene regulation verification of this invention demonstrates that the SST subtype interneurons in the lateral orbitofrontal cortex are the root cause of cognitive flexibility impairment in obsessive-compulsive disorder. Figure 6 This is a schematic diagram illustrating how the conditional gene regulation verification of this invention demonstrates that the PV subtype interneuron in the lateral orbitofrontal cortex is the root cause of the reduced cognitive flexibility in obsessive-compulsive disorder. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] The experimental subjects included Sapap3 gene knockout obsessive-compulsive disorder model mice, SST-Cre and PV-Cre tool mice, Sapap3 conditional knockout mice and conditional knock-in mice. All of these experimental mice were existing mature resources, which were obtained from the collaborating laboratory and bred and expanded in a standard SPF-level animal room.

[0020] I. Locating the time window of SST subtype interneuron dysfunction using fiber optic calcium imaging Sapap3+ / +:SST-Cre+ / - and Sapap3- / -:SST-Cre+ / - mice from the same littermate were selected and underwent stereotactic surgery at 3 months of age. After anesthesia, the mice were fixed in a stereotactic apparatus, with the anterior fontanelle as the origin, and the coordinates of the lateral orbitofrontal cortex (AP: +2.6 mm, ML: ±1.5 mm, DV: -1.6 mm) were determined using a brain atlas. 240 nL of AAV-DIO-GCaMP6m virus was injected at a rate of 60 nL / min using a microinjector to specifically label SST subtype interneurons (attached...). Figure 1 A). The needle was left in place for 10 minutes after injection, then slowly withdrawn, and an optical fiber and head fixation plate were implanted. Each mouse was housed individually after the surgery.

[0021] After 3-4 weeks of recovery, the mice were deprived of water for 36 hours, maintaining their weight at 85% of their pre-deprivation weight. Before each training session, the mice's heads were fixed in place, but they were still allowed to run on the wheel (see attached image). Figure 1 A). After fixing the mouse's head, allow the mouse to acclimatize for 1 hour.

[0022] After completing a 3-day adaptive training period, the mice can then proceed to the formal reverse learning task.

[0023] The entire reversal learning process is divided into the following 5 stages: odor discrimination (OD), discrimination reinforcement (DR), early reversal stage (ER), late reversal stage (LR), and reversal reinforcement (RR).

[0024] Odor Discrimination Learning (OD) Phase: At the start of each trial, mice are randomly exposed to one of two odors, odor 1 and odor 2 (each odor accounts for 50% of the trials), with each odor lasting for 1 second. During the 3-5 second time window following the introduction of odor 1, licking the spitting area earns a reward (hit). Not licking the spitting area or licking it outside the time window results in no reward (miss). Odor 2 is not associated with rewards. Licking the spitting area within the first 6 seconds after odor 2 is introduced is recorded as a false alarm; otherwise, it is a correct rejection. Each trial lasts 15 seconds. Every 20 trials constitute a session. The behavioral standard is an accuracy rate of over 80% for two consecutive sessions.

[0025] Odor Differentiation Reinforcement Phase (DR): After achieving the learning standard, 10 more sets of reinforcement learning will be conducted.

[0026] Reversal Phase: The dependency relationship between the two odors and the reward is reversed, that is, odor 2 can obtain a reward, while odor 1 does not. Furthermore, the reversal phase is divided into early reversal (ER) and late reversal (LR) based on the first time a 60% accuracy rate is achieved.

[0027] The Reverse Reinforcement (RR) phase is a reinforcement learning phase that follows the achievement of the learning standard, and involves 10 more sets of reinforcement learning.

[0028] Throughout the reverse learning process, GCaMP6m fluorescence signals were acquired using a fiber optic recording system. The dynamic changes in calcium signals in SST subtype interneurons were analyzed at different learning stages and for different behavioral outcome types.

[0029] The results showed that the activity of SST subtype interneurons in KO mice was significantly lower than that in WT mice during correct rejection and false alarm trials in the early stages of reversal, particularly during cue presentation and behavioral selection. (See attached image) Figure 1 B); Observation of interneuron activity in the SST subtype revealed that, in both WT and KO mice, under false alarm conditions, neuronal activity significantly increased after rule reversal, and gradually decreased with training (see appendix). Figure 1 (C and D).

[0030] Regression analysis of SST subtype interneurons and the accuracy of behavioral performance at specific stages showed a significant negative correlation between the two (see appendix). Figure 1 E).

[0031] II. Locating the time window of PV subtype interneuron dysfunction using fiber optic calcium imaging Sapap3+ / +:PV-Cre+ / - and Sapap3- / -:PV-Cre+ / - mice from the same littermate were selected and underwent stereotactic surgery at 3 months of age. After anesthesia, the mice were fixed in a stereotactic apparatus with the anterior fontanelle as the origin, and the lateral orbitofrontal cortex coordinates (AP: +2.6 mm, ML: ±1.5 mm, DV: -1.6 mm). 240 nL of AAV2 / 9-DIO-GCaMP6m virus was injected at a rate of 60 nL / min using a microinjector to specifically label PV subtype interneurons (attached...). Figure 2 A). After injection, the needle was left in place for 10 minutes, then slowly withdrawn, and an optical fiber and head fixation plate were implanted. Mice were housed individually post-surgery.

[0032] Two weeks after recovery, the mice were deprived of water for 36 hours to maintain their weight at 85% of their pre-deprivation weight. Before each training session, the mice's heads were fixed in place, yet they were still allowed to run on the wheel (see attached image). Figure 2 A). After fixing the mouse's head, allow the mouse to acclimatize for 1 hour.

[0033] After completing a 3-day adaptive training period, the mice can then proceed to the formal reverse learning task.

[0034] For PV subtype interneurons, KO mice showed significantly reduced activity in false alarm trials during both the early and late stages of reversal, continuing into the feedback period (see appendix). Figure 2 B).

[0035] Observation of interneuron activity in PV subtypes revealed that, in both WT and KO mice, under false alarm conditions, neuronal activity significantly increased after rule reversal, and gradually decreased with training (see appendix). Figure 2 (C and D).

[0036] Furthermore, the activity intensity of PV subtype interneurons was also significantly negatively correlated with the accuracy of behavioral performance at specific stages (see appendix). Figure 2 E).

[0037] III. Validating the causal relationship between SST subtype interneuron dysfunction and reverse learning disability through optogenetics Sapap3- / -:SST-Cre+ / - mice were injected bilaterally into the lateral orbitofrontal cortex with AAV-DIO-ChR2-EGFP virus, which specifically expresses the light-sensitive channel protein ChR2 in SST subtype interneurons; the control group was injected with the corresponding fluorescent protein control virus EGFP. After viral injection, optical fibers and head fixation plates were implanted bilaterally, and the mice were housed individually post-surgery. Reverse learning was assessed 3-4 weeks after full recovery.

[0038] Because the time window for the development of neurological dysfunction in SST subtype interneurons is early in the trial, near the cue firing phase, a stimulation time window of 0-3 seconds was set (see attached). Figure 3 A); Simultaneously, to further investigate whether stimulation within a time window when no functional abnormalities occurred in the SST subtype interneurons would affect the reversal learning performance in mice, a light modulation experiment with a 6-9 s time window was also set up (see appendix). Figure 3 B).

[0039] The results showed that during the reversal learning process, applying 473 nm blue light stimulation (5 ms pulse, 40 Hz, 5-10 mW) within the 0-3 s time window of trials with rewards before reversal but no rewards after reversal specifically activated SST subtype interneurons, significantly improving reversal learning impairment in KO mice. The number of learning attempts required to complete the entire reversal learning process was significantly reduced compared to the control group, primarily shortening the number of learning attempts in the early stages of reversal, with no significant effect on the later stages of reversal. (See appendix) Figure 3 CE).

[0040] The results showed that during the reversal learning process, applying 473 nm blue light stimulation (5 ms pulse, 40 Hz, 5~10 mW) within the 6-9 s time window of trials with rewards before reversal but no rewards after reversal did not significantly improve reversal learning impairment in KO mice by specifically activating SST subtype interneurons. The number of learning attempts required to complete the entire reversal learning process was comparable to that of the control group, and there was no significant effect on either the early or late stages of reversal. (See appendix) Figure 3 IK).

[0041] AAV-DIO-eNpHR3.0-mCherry virus was injected bilaterally into the lateral orbitofrontal cortex of wild-type SST-Cre+ / - mice. The experimental group mice showed specific expression of the light-sensitive chloride channel eNpHR3.0 in the SST subtype interneurons; the control group expressed the corresponding control fluorescent protein mCherry. Following virus injection, optical fibers were implanted bilaterally, and head fixation plates were attached. The mice were housed individually post-surgery and allowed 3-4 weeks of full recovery before undergoing a reverse learning test.

[0042] The results showed that during the reversal learning process, applying 589 nm yellow light stimulation (3-5 mW) within the 0-3 s time window of trials where there was a reward before reversal but no reward after reversal specifically inhibited SST subtype interneurons, significantly inducing reversal learning impairment in WT mice. The number of learning attempts required to complete the entire reversal learning process was significantly increased compared to the control group, primarily prolonging the learning attempts in the early stages of reversal, with no significant effect on the later stages of reversal. (See appendix) Figure 3 FH).

[0043] The results showed that during the reversal learning process, applying 589 nm yellow light stimulation (3-5 mW) within a 6-9 s time window to trials where there was a reward before reversal but no reward after reversal did not induce reversal learning impairment in WT mice by specifically inhibiting SST subtype interneurons. The number of learning attempts required to complete the entire reversal learning process was comparable to that of the control group, and there was no significant effect on the early or late stages of reversal. (See appendix) Figure 3 LN).

[0044] IV. Validating the causal relationship between PV subtype interneuron dysfunction and reverse learning disability through optogenetics Sapap3- / -:PV-Cre+ / - mice were injected bilaterally into the lateral orbitofrontal cortex with AAV-Flex-ChrimsonR-Tdtomato or AAV-DIO-ChR2-EGFP virus, which specifically expresses the light-sensitive proteins ChrimsonR or ChR2 in PV subtype interneurons. Following viral injection, fiber optic implantation and head fixation plates were performed, and mice were housed individually post-surgery. Reversal learning tests were conducted 3-4 weeks after full recovery.

[0045] Optogenetic regulation occurs during the inversion phase, with 638 nm or 473 nm light stimulation (5–10 mW) applied during time windows of 0–3 s and 6–9 s, respectively, specifically activating PV subtype interneurons (attached). Figure 4 A, B).

[0046] The results showed that after rule reversal, photoactivation of PV subtype interneurons within a 0-3s time window (previously rewarded but no longer rewarded) significantly improved reversal learning impairment and shortened the number of learning attempts in the early and late stages of reversal (see appendix). Figure 4 CE); while 6-9 s photoactivation of PV subtype interneurons can also significantly improve reversal learning impairment, but only shortens the number of learning sessions in the early stages of reversal, and has no effect on behavioral performance in the later stages of reversal (see appendix). Figure 4 IK).

[0047] Wild-type PV-Cre+ / - mice were injected with the same virus, which specifically expressed eNpHR3.0 in PV subtype interneurons. The control group was injected with mCherry control virus. After viral injection, fiber optic implantation and head fixation plates were attached, and mice were housed individually post-surgery. Reversal learning tests were performed 3-4 weeks after full recovery.

[0048] Similarly, in the regulation experiments in wild-type mice, regulation was also carried out within time windows of 0-3s and 6-9s. That is, after the rule was reversed, continuous illumination of 589 nm yellow light (3-5 mW) was applied within the time windows of 0-3s or 6-9s for trials that were previously rewarded but were no longer rewarded, specifically inhibiting the activity of PV subtype interneurons.

[0049] The results showed that inhibiting PV subtype interneurons within the 0-3s time window could induce reversal learning impairment and prolong the number of learning cycles in both the early and late stages of reversal (see appendix). Figure 4 FH). However, inhibiting PV subtype interneurons within the 6-9s time window only affects the number of learning iterations required in the early stages of inversion learning, and has no effect on the later stages (see appendix). Figure 4 LN).

[0050] The above-mentioned two-way validation results of optogenetic regulation prove (attached) Figure 3 and attached Figure 4 A causal relationship exists between interneuron dysfunction in the SST and PV subtypes of the lateral orbitofrontal cortex and reduced cognitive flexibility in obsessive-compulsive disorder.

[0051] V. Verification through conditional gene manipulation that SST subtype interneurons are the source of cognitive flexibility impairment in obsessive-compulsive disorder model mice. Three-month-old Sapap3 conditional knockout mice were stereotactically injected into the lateral orbitofrontal cortex with AAV-SST-Cre adeno-associated virus at a titer of approximately 2 × 10⁻⁶. 12 Vg / mL, 200 nL, specifically knocking out Sapap3 gene expression only in SST subtype interneurons. The control group received the same virus injected into the lateral orbitofrontal cortex of wild-type mice. Following viral injection, a head fixation plate was implanted. In some experiments, co-injection of the calcium-imaging virus enabled monitoring of SST subtype interneuron and pyramidal neuron activity during inversion learning (see appendix). Figure 5 A).

[0052] The results showed that specific knockout of Sapap3 expression in SST subtype interneurons induced inversion learning impairment, significantly increasing the number of learning iterations required to complete the entire inversion learning process compared to the control group. Furthermore, this only caused abnormalities in the early stages of inversion and had no effect on later stages. (See appendix) Figure 5 B).

[0053] Fiber optic recordings confirmed that knocking out the Sapap3 gene significantly reduced the activity of SST subtype interneurons under both false alarm and correct rejection conditions in the early stages of reversal (see appendix). Figure 5 C), while the activity of pyramidal neurons is significantly increased (see appendix). Figure 5 D).

[0054] In 3-month-old Sapap3 conditional knock-in mice, AAV-SST-Cre virus was injected into the lateral orbitofrontal cortex, specifically restoring the Sapap3 gene only in SST subtype interneurons. Control group knockout mice were injected with AAV-SST-Cre. Following virus injection, a head fixation plate was implanted. In some experiments, co-injection of the calcium imaging virus was used to monitor the activity of SST subtype interneurons and pyramidal neurons during inversion learning (see appendix). Figure 5 E).

[0055] The results showed that restoring the Sapap3 gene only in the interneurons of the SST subtype was sufficient to correct the inversion learning impairment, significantly reducing the number of learning iterations required to complete the entire inversion learning process compared to the control group, and only reducing the number of learning iterations in the early stages of inversion (see appendix). Figure 5 F).

[0056] Fiber optic recordings confirmed that, upon restoration of the Sapap3 gene, the activity of SST subtype interneurons was significantly increased under both false alarm and correct rejection conditions in the early stages of reversal (see appendix). Figure 5 G), while pyramidal neuron activity was significantly reduced (see appendix) Figure 5 H).

[0057] The above-mentioned gene-level verification from both positive and negative perspectives confirms, from the dimensions of sufficiency and necessity, that the selective loss of function of the Sapap3 interneuron in the SST subtype of the lateral orbitofrontal cortex leads to impaired ability to explore new rules in the early stages of rule reversal.

[0058] VI. Verification through conditional gene manipulation that PV subtype interneurons are the source of cognitive flexibility impairment in obsessive-compulsive disorder model mice. Three-month-old Sapap3 conditional knockout mice were stereotactically injected into the lateral orbitofrontal cortex with AAV-PV-Cre virus at a titer of approximately 2 × 10⁻⁶. 12 vg / mL, volume 200 nL, specifically knocking out Sapap3 gene expression only in PV subtype interneurons. The control group was injected with the same virus into the lateral orbitofrontal cortex of wild-type mice (see attached image). Figure 6 A).

[0059] The results showed that specific knockout of Sapap3 expression in PV subtype interneurons effectively induced reverse learning impairment, leading to abnormalities in both early and late learning stages (see appendix). Figure 6 B).

[0060] Fiber optic recordings confirmed that knocking out the Sapap3 gene significantly reduced the activity of PV subtype interneurons under false alarm conditions in both the early and late stages of reversal (see appendix). Figure 6 C), while the activity of pyramidal neurons is significantly increased (see appendix). Figure 6 D).

[0061] In 3-month-old Sapap3 conditional knock-in mice, AAV-PV-Cre virus was injected into the lateral orbitofrontal cortex, resulting in specific restoration of the Sapap3 gene only in PV subtype interneurons. Control group KO mice were injected with AAV-PV-Cre (see attached image). Figure 6 E).

[0062] The results showed that restoring the Sapap3 gene only in PV subtype interneurons was sufficient to correct reverse learning impairment, significantly reducing both early and late learning attempts (see appendix). Figure 6 F).

[0063] Fiber optic recordings confirmed that, upon restoration of the Sapap3 gene, the activity of PV subtype interneurons was significantly increased in the early and late stages of the reversal under false alarm conditions (see appendix). Figure 6 G), while pyramidal neuron activity was significantly reduced (see appendix) Figure 6 H).

[0064] The above-mentioned positive and negative gene-level verifications confirm, from the two dimensions of sufficiency and necessity, that the loss of function of the Sapap3 interneuron in the PV subtype of the lateral orbitofrontal cortex leads to impaired ability to explore and maintain new rules throughout the rule reversal process.

[0065] VII. Based on the above evidence, the cell-specific therapeutic target was finally determined. Based on the evidence from the three levels corresponding to the two types of neurons, this invention ultimately clarifies that the SST subtype interneurons and PV subtype interneurons in the lateral orbitofrontal cortex are cell-specific therapeutic targets for treating cognitive flexibility impairment related to obsessive-compulsive disorder. The SST subtype interneurons correspond to the function of exploring new rules in the early stage of rule reversal, while the PV subtype interneurons correspond to the function of exploring and maintaining new rules throughout the entire reversal process, and can be used as targets for precise intervention.

[0066] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder, characterized in that, Includes the following steps: First, fiber optic imaging was used to record the dynamic changes in neuronal activity of SST and PV subtype interneurons in the lateral orbitofrontal cortex of an obsessive-compulsive disorder (OCD) animal model and normal control animals during a reverse learning task, locating the time window and event type of functional abnormality. Based on the identified time window of functional abnormality, optogenetics was used to specifically regulate the activity of specific subtype interneurons to verify whether correcting or simulating functional abnormalities can improve or induce cognitive flexibility impairment, thereby establishing a causal relationship. Then, conditional gene manipulation was used to specifically knock out or restore the Sapap3 gene in specific subtype interneurons of the lateral orbitofrontal cortex of adult animals, verifying that functional abnormalities of specific subtype interneurons in specific brain regions are the root cause of cognitive flexibility impairment, and identifying the cellular targets for treating cognitive flexibility reduction in OCD, namely, SST and PV subtype interneurons in the lateral orbitofrontal cortex.

2. The cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder according to claim 1, characterized in that: The location of the time window and event type of the functional abnormality includes: for SST subtype interneurons, in the early stage of reversal learning, during the presentation of cues that were rewarded before reversal but not after reversal and the behavioral selection stage, neuronal activity is abnormally reduced; for PV subtype interneurons, in the early and late stages of reversal learning, during the presentation of cues that were incorrectly selected after the presentation of unrewarded odors and the result feedback stage, neuronal activity is abnormally reduced.

3. The cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder according to claim 1, characterized in that: The specific regulation of the optogenetic technology includes: for SST subtype interneurons, specific regulation is performed on the 0-3 second cue presentation phase after rule reversal, where there is a reward before reversal but no reward after reversal; for PV subtype interneurons, specific regulation is performed on the 0-3 second cue presentation phase and the 6-9 second result feedback phase in trials without reward after rule reversal.

4. The cellular target for treating cognitive flexibility impairment in obsessive-compulsive disorder according to claim 1, characterized in that: The conditional gene manipulation technology includes: specifically knocking out the Sapap3 gene in interneurons of the SST or PV subtypes of the lateral orbitofrontal cortex of adult mice, and observing whether it induces cognitive flexibility impairment; then specifically re-expressing the Sapap3 gene in interneurons of the SST or PV subtypes of the lateral orbitofrontal cortex of adult mice, and observing whether it salvages cognitive flexibility impairment.

5. The use of a cellular target for treating cognitive flexibility impairment according to any one of claims 1-4 in the preparation of drugs or interventions for treating cognitive flexibility disorders.

6. The application according to claim 5, characterized in that: The application of the SST subtype interneurons is as follows: to treat cognitive flexibility impairment, which is mainly manifested by impaired ability to explore new rules, with the intervention time window being the early stage of rule reversal.

7. The application according to claim 5, characterized in that: The application of the PV subtype interneurons is to treat cognitive flexibility impairment that simultaneously impairs the ability to explore new rules and maintain new rules, with the intervention time window covering the early and late stages of rule reversal.

8. The application according to claim 5, characterized in that: The method involves restoring cognitive flexibility through targeted intervention of specific inhibitory interneuron subclasses in the lateral orbitofrontal cortex, and the intervention includes optogenetic regulation and gene therapy.

9. The application according to claim 8, characterized in that: The targeted intervention is implemented in adulthood and is not limited to critical developmental periods.

Images