Application of cytokine interleukin-17D and histone milk acylation and acetylation modification induced by cytokine interleukin-17D in treatment of psoriasis recurrence

Abstract

The invention discloses a cytokine interleukin-17D (IL-17D) and application of histone milk acylation and acetylation modification induced by the cytokine interleukin-17D (IL-17D) as drug targets in preparation of drugs for treating psoriasis recurrence. In the pathopoiesis process of psoriasis, IL-17D promotes the formation of inflammatory memory of keratinocytes by activating the receptor CD93 of IL-17D to promote the modifications such as histone acetylation and lactoacylation, so that the recurrence of psoriasis is aggravated. After an Il17d gene of a mouse and an IL-17D receptor gene Cd93 in keratinocytes are knocked out, histone modification in psoriasis recurrence skin lesion and expression of psoriasis-related pathogenic cytokines and chemotactic factors are reduced, and scurf accumulation and epidermal thickness in the skin lesion during psoriasis recurrence are obviously reduced; the relapse of the psoriasis can be obviously inhibited after a histone acetyl and lactoyltransferase P300 / CBP inhibitor A-485 is injected into a mouse with the psoriasis to inhibit IL-17D induced histone acetylation and lactoacylation modification.

Description

Technical Field

[0001] The present invention relates to the fields of biochemistry, molecular biology, immunology and dermatology, and relates to the application of cytokine interleukin-17D and histone lactylation and acetylation modifications induced by the cytokine interleukin-17D in the treatment of relapse of psoriasis. Background Art

[0002] Although a variety of biological agents or topical medications can successfully treat psoriasis, psoriasis often recurs in the original lesions after the patient stops using the medication for a period of time. According to statistics, the probability of psoriasis recurrence is nearly 100%, of which the probability of in situ recurrence is 90%. 1 Psoriasis is often accompanied by complications such as diabetes, arthritis and cardiovascular disease, which not only reduces the patient's quality of life but also threatens the patient's life and health. 2,3 Therefore, in-depth exploration of the molecular mechanism of psoriasis relapse will provide new methods and new targets for the treatment of psoriasis relapse.

[0003] The TNFα-IL23-IL17A axis plays an important role in the occurrence and development of psoriasis. Neutralizing antibodies developed against these cytokines can effectively treat psoriasis. However, these neutralizing antibody therapies can only inhibit the production and function of inflammatory factors, and cannot eliminate cells related to the pathogenesis of psoriasis. Therefore, once the drug is stopped, pathogenic cells will continue to secrete cytokines and chemokines to induce the recurrence of psoriasis. Currently, memory T cells (Tissue-resident memory T cells, T RM ) is considered the main cause of psoriasis relapse 4,5 A large number of T cells can be detected in the skin after psoriasis has subsided. RM cells, which can express the pathogenic factors IL-22 and IL-17A, thereby activating keratinocytes and possibly inducing the recurrence of psoriasis 4,5 But T RM Can migrate to other parts of the skin or other tissues away from the skin through lymphatic and blood circulation 6,7 Therefore, T RM It may not be the only cause of psoriasis in situ recurrence.

[0004] In addition to T RM In addition, keratinocytes in the basal layer of the epidermis form inflammatory memory in the psoriasis inflammatory microenvironment and enhance the inflammatory response ability of keratinocytes. 8 In addition, keratinocytes, as structural cells of the epidermis, do not migrate with the blood and lymphatic circulation. Therefore, the inflammatory response and inflammatory memory of keratinocytes may be the cause of the recurrence of psoriasis in situ. 8,9It is known that histone modification-mediated increase in chromatin accessibility and the continuous binding of transcription factor JUN at chromatin openings are key to the formation and maintenance of keratinocyte "inflammatory memory" 10 The inflammatory microenvironment can promote the increase of H3K4 methylation (H3K4me1 / 3) and acetylation (H3K27ac) in keratinocytes 10 Among them, H3K4me1 / 3 recruits chromatin remodeling protein (Chromodomain-Helicase DNA-binding 1, CHD1) to promote chromatin accessibility at the transcription start site (TSS). 11,12 The negative charge carried by H3K27ac leads to increased chromatin accessibility by repelling each other or negatively charged DNA. 13,14 In the pathogenesis of psoriasis, activation of STAT3 recruits constitutive transcription factors FOS and JUN to form a transcription factor complex that enters the loose chromatin site and promotes gene transcription. 10 After inflammation subsides, H3K4me1, along with low levels of H3K27ac and the transcription factor JUN, can maintain chromatin openness, making it readily accessible, marking the formation of "inflammatory memory." 10 .

[0005] Although it is known that keratinocytes can form inflammatory memory in the inflammatory microenvironment of psoriatic lesions, it is not clear which factors can promote histone modifications in keratinocytes to induce the formation of inflammatory memory. It is known that changes in cell metabolism are the driving force for the formation of histone modifications in innate immune memory. 15 For example, the glutamine hydrolysis pathway hydrolyzes extracellular glutamine into succinate in the TCA cycle, and the accumulated succinate will be further converted into fumarate. 16 Fumaric acid can inhibit the activity of histone demethylase KDM5, thereby upregulating the level of H3K4me3 in cells 17 Acetyl-CoA, as a substrate for histone acetylation, can promote the formation of H3K27ac. 16,18 Therefore, finding the factors that induce inflammatory memory formation of keratinocytes in the psoriasis inflammatory environment and exploring its regulatory mechanism are the key to the clinical treatment of recurrent skin diseases such as psoriasis relapse. Summary of the Invention

[0006] The present invention addresses the deficiencies of the above-mentioned prior art and proposes the use of the cytokine interleukin-17D (IL-17D) and its induced histone acetylation and lactylation modifications as targets for the preparation of drugs or preparations for the prevention and / or treatment of psoriasis relapse. The present invention proposes novel methods and strategies for preventing and / or treating psoriasis relapse by targeting IL-17D or its induced histone acetylation and lactylation, effectively and specifically inhibiting the inflammatory memory of keratinocytes in psoriasis, thereby suppressing the inflammatory response of keratinocytes during psoriasis relapse, and thereby delaying or inhibiting the recurrence of psoriasis.

[0007] During the pathogenesis of psoriasis, IL-17D can induce modifications such as histone acetylation (H3K27ac) and lactylation (H3K18la) in keratinocytes, causing keratinocytes to acquire inflammatory memory, thereby aggravating the recurrence of psoriasis. The present invention knocks out the Il17d gene or the Cd93 gene in keratinocytes in mice. Histone modifications in recurrent psoriasis lesions, the expression of psoriasis-related pathogenic cytokines and chemokines, and the accumulation of psoriasis and epidermal thickness in the lesions are significantly reduced during psoriasis relapse. Injecting the histone acetyltransferase P300 / CBP inhibitor A-485 into psoriasis mice to inhibit IL-17D-induced histone acetylation and lactylation modifications can significantly inhibit the recurrence of psoriasis. The present invention establishes a psoriasis relapse model in Il17d-deficient mice, which significantly reduces or inhibits psoriasis accumulation, ear thickness, epidermal thickness, H3K27ac and H3K18la levels, and the expression of pathogenic cytokines and chemokines in relapsed psoriasis lesions. Injection of IL-17D protein significantly reverses psoriasis relapse and H3K27ac and H3K18la levels in lesional skin in Il17d-deficient mice. Therefore, the present invention proposes that IL-17D and its induced histone acetylation and lactoylation modifications may serve as therapeutic targets for delaying psoriasis relapse.

[0008] The term "IL-17D" described in the present invention refers to the cytokine interleukin-17D, which belongs to the IL-17 family of cytokines and is a member of the IL-17 family. It can activate its receptor CD93 to perform its biological function.

[0009] The present invention provides the feasibility of using IL-17D and its induced histone acetylation and lactoylation modifications as targets for delaying the recurrence of psoriasis.

[0010] The present invention provides the use of cytokine interleukin-17D and histone lactylation and acetylation modifications induced by the cytokine interleukin-17D as drug targets in the preparation of drugs for preventing and / or treating the recurrence of psoriasis.

[0011] The cytokine interleukin-17D promotes histone H3K18 lactylation (H3K18la) and H3K27 acetylation (H3K27ac) modifications in keratinocytes by activating the receptor CD93 on keratinocytes, thereby promoting keratinocytes to form inflammatory memory to express more cytokines and chemokines, thereby promoting the recurrence of psoriasis; therefore, the purpose of inhibiting the recurrence of psoriasis can be achieved by inhibiting the activity of the cytokine interleukin-17D in cells or tissues.

[0012] The method for inhibiting the activity of the cytokine interleukin-17D in cells or tissues comprises knocking down the receptor CD93 of the cytokine interleukin-17D on keratinocytes using CD93 siRNA.

[0013] The psoriasis includes but is not limited to vulgaris, pustular, guttate psoriasis and the like.

[0014] The IL-17D induces keratinocytes to acquire inflammatory memory and enhances the expression of cytokines and chemokines when keratinocytes are subjected to secondary stimulation from the outside world such as poly(I:C) (polyinosinic acid, simulating dsRNA released by damaged cells).

[0015] The cytokines include but are not limited to IL36G, IL18, and IL33, and the chemokines include but are not limited to CXCL1 and CCL20.

[0016] The IL-17D-induced H3K27ac and H3K18la are involved in the inflammatory memory of keratinocytes.

[0017] The present invention also provides that the loss of IL-17D in mice reduces or inhibits the levels of H3K27ac, H3K18la and the expression of Il17a, Il23, Cxcl1 and Ccl20 in skin lesions, thereby delaying the recurrence of psoriasis.

[0018] The present invention also provides that the specific deletion of the IL-17D receptor CD93 in mouse keratinocytes will reduce or inhibit the levels of H3K27ac, H3K18la and the expression of Il17a, Il23, Cxcl1 and Ccl20 in skin lesions, thereby delaying the recurrence of psoriasis.

[0019] The present invention also provides that the IL-17D-induced histone H3K27ac, H3K18la and other modifications can be inhibited by the histone acetyltransferase P300 / CBP inhibitor A-485, causing keratinocytes to lose inflammatory memory, thereby reducing or inhibiting the expression of Il17a, Il23, Cxcl1 and Ccl20, and thereby inhibiting the recurrence of psoriasis.

[0020] The present invention also provides an inhibitor of histone lactylation and acetylation modification induced by cytokine interleukin-17D, wherein the inhibitor of histone lactylation and acetylation modification includes histone acetyltransferase P300 / CBP inhibitor A-485 and the like.

[0021] The histone acylation and acetylation modifications include H3K27ac, H3K18la, etc.

[0022] The present invention also provides the use of the inhibitor of histone lactylation and acetylation modification induced by the cytokine interleukin-17D in the preparation of a drug for preventing and / or treating the recurrence of psoriasis.

[0023] In the application of the present invention, the psoriasis includes but is not limited to vulgaris, pustular, and guttate psoriasis.

[0024] In the application of the present invention, the inhibitor can be used alone or in combination with other drugs.

[0025] The present invention also provides a drug / pharmaceutical composition comprising the CD93 siRNA and an inhibitor as described above.

[0026] The present invention also provides the use of the medicine / drug composition in preparing a medicine for preventing and / or treating the recurrence of psoriasis.

[0027] In the application of the present invention, the drug / drug composition can be used alone or in combination with other drugs.

[0028] The present invention also provides a detection reagent / kit, which can be used to detect the expression of IL-17D as described above.

[0029] The present invention also provides the use of the detection reagent / kit in preparing a drug for preventing and / or treating the recurrence of psoriasis.

[0030] In the application of the present invention, the psoriasis includes but is not limited to vulgaris, pustular, and guttate psoriasis.

[0031] In the application of the present invention, the detection reagent / kit can be used alone or in combination with other drugs.

[0032] The present invention also provides a method for screening candidate drugs for preventing and / or treating the recurrence of psoriasis, the method comprising detecting the effect of the candidate drug on the biological effects of IL-17D as described above and its induced H3K27ac and H3K18la in a subject or a sample obtained from the subject.

[0033] Among them, after using the candidate drug, the biological function of IL-17D inducing H3K27ac and H3K18la is inhibited, indicating that the candidate drug has the effect of preventing and / or treating the recurrence of psoriasis; the improvement of the biological function of IL-17D includes an increase in the expression level of IL-17D.

[0034] The present invention also provides a method for preventing and / or treating the recurrence of psoriasis in a subject in need thereof, or a method for assessing the severity of recurrence in a psoriasis patient, or a method for predicting or prognosticating whether the patient is suitable for treatment targeting IL-17D or H3K27ac, H3K18la, the method comprising: determining the level of IL-17D or H3K27ac, H3K18la in a psoriasis tissue sample from the patient or subject, wherein the level is used to prevent or treat the recurrence of psoriasis in the subject, or to assess the severity of psoriasis recurrence in a patient, or to predict or prognosticate whether the patient is suitable for treatment targeting IL-17D or H3K27ac, H3K18la.

[0035] In the above-mentioned application and method of the present invention, the psoriasis includes but is not limited to vulgaris, pustular, and guttate psoriasis.

[0036] The beneficial effects of the present invention include: by identifying IL-17D in the psoriasis inflammatory environment as a factor involved in psoriasis relapse, the present invention proposes that IL-17D activates its receptor CD93 to induce H3K27ac and H3K18la in keratinocytes, training keratinocytes to acquire inflammatory memory, and enhancing the keratinocytes' expression of more and faster cytokines and chemokines upon secondary stimulation, thereby enhancing the keratinocytes' inflammatory response capacity and promoting the recurrence of psoriasis. The present invention brings new hope for the clinical treatment or prevention of recurrent skin diseases such as psoriasis relapse and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without paying any creative work.

[0038] Figure 1Figure 1 shows IL-17D-induced histone modifications and inflammatory memory in keratinocytes. Figure A shows the levels of histone H3K27ac and H3K18la in keratinocytes after IL-17D treatment. Figure B shows the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20 after IL-17D pretreatment and subsequent poly(I:C) treatment.

[0039] Figure 2 IL-17D deficiency delays the relapse of psoriasis in mice. Panel A shows photographs of WT and Il17d-deficient mice during the uninduced psoriasis phase (also known as the "normal phase"), the active phase (also known as the "active phase"), and the relapse phase (also known as the "relapse phase"). Panel B shows ear thickness in WT and Il17d-deficient mice during the uninduced psoriasis phase (also known as the "normal phase"), the active phase (also known as the "active phase"), and the relapse phase (also known as the "relapse phase"). Panel C shows epidermal thickness in lesions in WT and Il17d-deficient mice during the uninduced psoriasis phase (also known as the "normal phase"), the active phase (also known as the "active phase"), and the relapse phase (also known as the "relapse phase"). Panel D shows histone modifications in lesions in WT and Il17d-deficient mice during the uninduced psoriasis phase (also known as the "normal phase"), the active phase (also known as the "active phase"), and the relapse phase (also known as the "relapse phase"), as detected by western blot. Figure E shows the ELISA detection of the expression of Il17a, Il23, Cxcl1 and Ccl20 in the skin lesions of WT and Il17d-deficient mice in the psoriasis uninduced stage (also known as the "normal stage"), psoriasis active stage and psoriasis relapse stage.

[0040] Figure 3 Injection of IL-17D protein exacerbates psoriasis relapse in Il17d-deficient mice. Figure A shows photographs of psoriasis lesions in Il17d-deficient mice during relapse after IL-17D protein supplementation. Figure B shows ear thickness in Il17d-deficient mice during relapse after IL-17D protein supplementation. Figure C shows epidermal thickness in psoriasis lesions in Il17d-deficient mice during relapse after IL-17D protein supplementation. Figure D shows western blot analysis of histone modification levels in psoriasis lesions in Il17d-deficient mice during relapse after IL-17D protein supplementation. Figure E shows ELISA analysis of Il17a, Il23, Cxcl1, and Ccl20 expression in psoriasis lesions in Il17d-deficient mice during relapse after IL-17D protein supplementation.

[0041] Figure 4The results indicate that specific knockout of Cd93 in keratinocytes delays the relapse of psoriasis in mice. Figure A shows photographs of WT and Il17d-deficient mice during the active phase of psoriasis (also referred to as the "active phase") and the relapse phase of psoriasis (also referred to as the "relapse phase"). Figure B shows ear thickness in WT and Il17d-deficient mice during the active phase of psoriasis (also referred to as the "active phase") and the relapse phase of psoriasis (also referred to as the "relapse phase"). Figure C shows epidermal thickness in lesions in WT and Il17d-deficient mice during the active phase of psoriasis (also referred to as the "active phase") and the relapse phase of psoriasis (also referred to as the "relapse phase"). Figure D shows histone modifications in lesions in WT and Il17d-deficient mice during the active phase of psoriasis (also referred to as the "active phase") and the relapse phase of psoriasis (also referred to as the "relapse phase") detected by western blot. Figure E shows the expression of Il17a, Il23, Cxcl1 and Ccl20 in the skin lesions of WT and Il17d-deficient mice during the active stage of psoriasis (also known as the "active stage") and the relapse stage of psoriasis (also known as the "relapse stage") detected by ELISA.

[0042] Figure 5 This figure shows that knocking down CD93 expression in keratinocytes using CD93 siRNA inhibits H3K18la and H3K27ac in keratinocytes, as well as inflammatory memory in keratinocytes. Figure A shows that knocking down CD93 in keratinocytes can inhibit IL-17D-induced H3K27ac and H3K18la modifications in keratinocytes, while lactate supplementation can significantly restore H3K27ac and H3K18la modifications. Figure B shows that knocking down CD93 in keratinocytes can inhibit IL-17D-induced inflammatory memory keratinocytes mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20, while lactate supplementation can significantly restore IL36G, IL18, IL33, CXCL1, and CCL20 mRNA expression in keratinocytes.

[0043] Figure 6 This study demonstrates that IL-17D-induced inflammatory memory in keratinocytes is dependent on histone acetylation and lactoylation modifications. Figure A shows that A-485 inhibits IL-17D-induced H3K27ac and H3K18la modifications in keratinocytes; Figure B shows that A-485 inhibits the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20 in IL-17D-induced inflammatory memory keratinocytes.

[0044] Figure 7A-485 injection delays psoriasis relapse in mice. Figure A shows a photograph of psoriasis lesions in WT mice at the relapse stage after A-485 injection; Figure B shows ear thickness in WT mice at the relapse stage after A-485 injection; Figure C shows epidermal thickness in psoriasis lesions in WT mice at the relapse stage after A-485 injection; Figure D shows histone modification levels in psoriasis lesions in WT mice at the relapse stage after A-485 injection by western blot; Figure E shows ELISA analysis of Il17a, Il23, Cxcl1, and Ccl20 expression in psoriasis lesions in WT mice at the relapse stage after A-485 injection. DETAILED DESCRIPTION

[0045] The present invention is further described in detail with reference to the following embodiments and accompanying drawings. The protection content of the present invention is not limited to the following embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the appended claims. The process, conditions, reagents, experimental methods, etc. for implementing the present invention, except for the contents specifically mentioned below, are common knowledge and common common sense in the art and are not particularly limited by the present invention.

[0046] Obviously, the described embodiments are only some of the embodiments of the present invention, rather than all of them. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention, its application, or use. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are intended to fall within the scope of protection of the present invention.

[0047] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not limiting. Therefore, other examples of the exemplary embodiments may have different values.

[0048] Unless otherwise specified, the test materials used in the examples are all conventional biochemical reagents.

[0049] The present invention discloses the use of the cytokine interleukin-17D (IL-17D) and its induced histone lactylation and acetylation modifications as drug targets in the preparation of drugs for treating the recurrence of psoriasis. During the pathogenic process of psoriasis, IL-17D induces the formation of inflammatory memory in keratinocytes by promoting histone acetylation, lactylation and other modifications, thereby aggravating the recurrence of psoriasis. After the present invention knocks out the Il17d gene in mice, histone modifications in the recurrent psoriasis lesions and the expression of psoriasis-related pathogenic cytokines and chemokines decrease, and the psoriasis accumulation and epidermal thickness in the lesions are significantly reduced when psoriasis relapses; after the histone acetyl and lactyl transferase inhibitor A-485 is injected into psoriasis mice to inhibit IL-17D-induced histone acetylation and lactylation modifications, the recurrence of psoriasis can be significantly inhibited. Therefore, the present invention discloses that IL-17D and its induced histone lactylation and acetylation modifications can be used as targets for preventing and treating the recurrence of psoriasis.

[0050] Example 1: IL-17D induces inflammatory memory in keratinocytes

[0051] After keratinocytes were stimulated with 0, 0.46, 0.911.38, 1.84 and 2.30nM IL-17D for 12 hours, western blot was used to detect the levels of H3K27ac and H3K18la in the lesions. After training keratinocytes with 60μg / mL IL-17D to form inflammatory memory, they were passaged. After passage to 1, 3, 5 and 7 days, 10μg / mL poly(I:C) was used to stimulate for 24 hours, and RNA was collected for RT-PCR to detect the mRNA expression of IL36G, IL18, IL33, CXCL1 and CCL20. The test found that IL-17D training can significantly upregulate the mRNA expression of IL36G, IL18, IL33, CXCL1 and CCL20 during poly(I:C) stimulation (such as Figure 1 B).

[0052] Example 2: IL-17D deficiency delays relapse of psoriasis in mice

[0053] A psoriasis model was induced by applying 25 mg of 5% imiquimod ointment (Imiquimod, IMQ) to each ear of 7-8 week-old wild-type mice and Il17d-deficient mice. 20 days after the inflammation subsided, IMQ was used again to induce psoriasis relapse. During the experiment, photos of the lesions were recorded with a camera, and the thickness of the mouse ears was recorded with a vernier caliper. Lesions were collected before psoriasis induction and during the active, resolving and relapse stages of psoriasis for paraffin embedding and H&E staining. Western blot was used to detect the levels of H3K27ac and H3K18la in the lesions, and ELISA was used to detect the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions. The test results showed that the absence of Il17d resulted in a decrease in psoriasis accumulation in the relapsed lesions (such as Figure 2 A), the thickness of the mouse ear becomes thinner (as shown in Figure 2 B), the epidermal thickness of the lesions is reduced (as shown in Figure 2 C), the levels of H3K27ac and H3K18la were reduced in the lesions (as shown in Figure 2 D), the expression of Il17a, Il23, Cxcl1, and Ccl20 was reduced in the lesions (as shown in Figure 2 E).

[0054] Example 3: IL-17D protein injection aggravates psoriasis relapse in Il17d-deficient mice

[0055] One day before induction, IL-17D or PBS was injected intradermally into the ears of 7- to 8-week-old Il17d-deficient mice, while PBS was injected into the ears of WT mice as a control. 25 mg of 5% imiquimod ointment (Imiquimod, IMQ) was applied to each ear to induce a psoriasis model, and 20 days after the inflammation subsided, IMQ was used again to induce psoriasis relapse. During the experiment, photos of the lesions were recorded using a camera, and the thickness of the mouse ears was recorded using a vernier caliper. After the relapse period, the lesions were collected for paraffin embedding and H&E staining, and the levels of H3K27ac and H3K18la in the lesions were detected by western blot, and the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions were detected by ELISA. The test results showed that the accumulation of psoriasis in the lesions of Il17d-deficient mice increased significantly after IL-17D was replenished (such as Figure 3 A), the thickness of the mouse ears was significantly restored (as shown in Figure 3 B), the epidermal thickness of the lesions recovered significantly (as shown in Figure 3 C), the levels of H3K27ac and H3K18la in the lesions were significantly restored (as shown in Figure 3 D), the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions was significantly restored (as shown in Figure 3E).

[0056] Example 4: Specific knockout of Cd93 in keratinocytes delays recurrence of psoriasis in mice

[0057] CD93 fl / fl Mice (control group) and K14 cre CD93 fl / fl A psoriasis model was induced by applying 25 mg of 5% imiquimod ointment (Imiquimod, IMQ) to each ear of mice (Cd93 specific knockout group in keratinocytes), and 20 days after the inflammation subsided, IMQ was used again to induce psoriasis relapse. During the experiment, photos of the lesions were recorded using a camera, and the thickness of the mouse ears was recorded using a vernier caliper. After the relapse period, the lesions were collected for paraffin embedding and H&E staining, and western blot was used to detect the levels of H3K27ac and H3K18la in the lesions, and ELISA was used to detect the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions. The test results showed that the accumulation of psoriasis in the lesions of mice with specific knockout of Cd93 in keratinocytes was significantly reduced (such as Figure 4 A), the thickness of the mouse ears decreased significantly (as Figure 4 B), the thickness of the epidermis of the lesions decreased significantly (as shown in Figure 4 C), the levels of H3K27ac and H3K18la in the lesions were significantly reduced (as shown in Figure 4 D), the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions was significantly decreased (as shown in Figure 4 E).

[0058] The above results indicate that IL-17D participates in and aggravates the recurrence of psoriasis in vivo and can serve as a target for preventing or treating the recurrence of psoriasis.

[0059] Example 5: Knockdown of CD93 in keratinocytes inhibits IL-17D-induced keratinocyte inflammatory memory

[0060] After knocking down CD93 expression in keratinocytes using CD93 siRNA, the cells were stimulated with 60 μg / mL IL-17D or 20 mM lactate for 12 hours before passage. On the third day of passage, cells were harvested and used for western blot analysis of H3K27ac and H3K18la levels. The results showed that knocking down CD93 significantly inhibited IL-17D-induced H3K27ac and H3K18la levels, while lactate supplementation significantly restored H3K27ac and H3K18la levels in keratinocytes (e.g. Figure 5At the same time, 10 μg / mL poly(I:C) was used to stimulate keratinocytes stimulated with IL-17D for 24 hours, and RNA was collected for RT-PCR to detect the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20. The test results showed that knockdown of CD93 significantly inhibited the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20 in inflammatory memory keratinocytes induced by IL-17D, and lactate supplementation could significantly restore the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20 in keratinocytes (as shown in Figure 2A). Figure 5 B).

[0061] Example 6: A-485 inhibits IL-17D-induced keratinocyte inflammatory memory

[0062] Keratinocytes were stimulated with 10 μM A-485 and 60 μg / mL IL-17D for 12 hours and then passaged. On the third day of passage, cells were harvested and used for western blot analysis of H3K27ac and H3K18la levels. The results showed that A-485 inhibited IL-17D-induced H3K27ac and H3K18la levels (e.g., Figure 6 A). IL-17D-stimulated keratinocytes were stimulated with 10 μg / mL poly(I:C) for 24 h, and RNA was collected for RT-PCR to detect the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20. The test results showed that A-485 inhibited the mRNA expression of IL36G, IL18, IL33, CXCL1, and CCL20 in IL-17D-induced inflammatory memory keratinocytes (as shown in Figure 6 B).

[0063] Example 7: A-485 injection delays relapse of psoriasis in mice

[0064] One day before induction, 10 μl of 120 μM P300 inhibitor (A-458) was injected intradermally into the ears of 7- to 8-week-old wild-type mice. At the same time, the corresponding solvent (DMSO) was injected into the ears of the control group of WT mice as a control. 25 mg of 5% imiquimod ointment (Imiquimod, IMQ) was applied to each ear to induce a psoriasis model, and 20 days after the inflammation subsided, IMQ was used again to induce psoriasis relapse. During the experiment, photos of the lesions were recorded using a camera, and the thickness of the mouse ears was recorded using a vernier caliper. After the relapse period, the lesions were collected for paraffin embedding and H&E staining. Western blot was used to detect the levels of H3K27ac and H3K18la in the lesions, and ELISA was used to detect the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions. The test results showed that the accumulation of psoriasis in the lesions of mice was significantly reduced after the injection of the P300 inhibitor (A-458). Figure 7 A), the thickness of the mouse ears was significantly reduced (as Figure 7 B), the thickness of the epidermis of the lesions was significantly reduced (as shown in Figure 7 C), the levels of H3K27ac and H3K18la in the lesions were significantly decreased (as shown in Figure 7 D), the expression of Il17a, Il23, Cxcl1 and Ccl20 in the lesions was significantly decreased (as shown in Figure 7 E).

[0065] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0066] As used in the present invention, the terms "include" and "comprising" are open expressions, that is, including the contents specified in the present invention, but not excluding other aspects.

[0067] As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0068] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, any changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the claims.

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Claims

1. Application of the cytokine interleukin-17D and its induced histone lactylation and acetylation modifications as drug targets in the preparation of drugs for preventing and / or treating the recurrence of psoriasis.

2. The use according to claim 1, characterized in that The cytokine interleukin-17D promotes histone H3K18 lactylation (H3K18la) and H3K27 acetylation (H3K27ac) modifications in keratinocytes by activating its receptor CD93, thereby promoting keratinocytes to form inflammatory memory to express more cytokines and chemokines, thereby promoting the recurrence of psoriasis.

3. The use according to claim 2, characterized in that The method for inhibiting the activity of the cytokine interleukin-17D in cells or tissues comprises knocking down the receptor CD93 of the cytokine interleukin-17D on keratinocytes using CD93 siRNA.

4. The use according to claim 2, characterized in that The histone H3K27ac and H3K18la modifications induced by the cytokine interleukin-17D can be inhibited by the histone acetyltransferase P300 / CBP inhibitor A-485, thereby causing keratinocytes to lose their inflammatory memory and inhibiting the recurrence of psoriasis.

5. A detection reagent / kit, characterized in that: It can be used to detect the expression of the cytokine interleukin-17D as claimed in claim 1.

6. Use of the detection reagent / kit according to claim 5 in the preparation of a medicament for preventing and / or treating the recurrence of psoriasis.

7. A method for screening candidate drugs for preventing and / or treating relapse of psoriasis, characterized in that: The method comprises detecting the effect of the candidate drug on the biological effect of the cytokine interleukin-17D as described in claim 1 in a subject or a sample obtained from a subject.

8. A method for preventing and / or treating the relapse of psoriasis in a subject in need thereof, or a method for assessing the severity of relapse in a psoriasis patient, or a method for predicting or prognosticating whether the patient is suitable for treatment targeting the cytokine interleukin-17D or H3K27ac, H3K18la, characterized in that: The method comprises: determining the level of the cytokine interleukin-17D or the H3K27ac, H3K18la in a psoriasis tissue sample from the patient or subject, wherein the level is used to prevent or treat the recurrence of psoriasis in the subject, or to assess the severity of psoriasis relapse in patients, or to predict or prognose whether the patient is suitable for treatment targeting the cytokine interleukin-17D or the H3K27ac, H3K18la.

9. The use according to any one of claims 1 to 4, the use according to claim 6, the method according to claim 7, and the method according to claim 8, characterized in that: The psoriasis includes vulgaris, pustular and guttate psoriasis.

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