CN-121986756-A - Construction method and application of ejection fraction retention type heart failure animal model

Abstract

The invention combines atrial fibrillation with hypertension, and develops a novel HFpEF double-beating animal model. In particular, the animal model of the present invention can develop stably to mild structural and functional disease stages within three weeks of double-stroke and reach end-stage and die gradually within the next two weeks, which is highly coincident with the existing human HFpEF pathophysiological phenotype. The animal model of the invention is stable and rapid in development in a staged mode, is easy to implement, and provides a valuable tool for researching the molecular mechanism of occurrence and development of diseases and developing medicines.

Inventors

• SHI HONGJUN
• LIN JIAYI

Assignees

• 西湖大学

Dates

Publication Date

20260508

Application Date

20241106

Claims (10)

1. 1. A method of constructing an animal model for heart failure with preserved ejection fraction, comprising the steps of: (a) Providing an animal subject, and (B) Atrial fibrillation and hypertension are caused to the individual animals.
2. 2. The method of claim 1, wherein the animal subject is a non-human mammal, and the non-human mammal comprises a bovine, ovine, canine, porcine, rabbit, non-human primate, rodent.
3. 3. The method of claim 1, wherein the method of causing atrial fibrillation comprises down-regulating Lkb gene expression in the atrium.
4. 4. The method of claim 3, wherein down-regulating comprises gene knockout or gene knockdown.
5. 5. The method of claim 3, wherein the method of down-regulating the expression of Lkb gene is selected from the group consisting of: (A1) Introducing a deletion mutation or a frame shift mutation into Lkb gene; (A2) Carrying out gene editing on Lkb gene; (A3) RNAi interference is carried out on Lkb gene; (A4) Any combination of the A1 to A3.
6. 6. The method of claim 1, wherein the method of causing hypertension comprises administering a boosting drug, formulation or composition to the animal subject.
7. 7. A kit is characterized in that, the kit comprises: (I) Gene editing agents responsible for atrial fibrillation, and (II) a medicament or pharmaceutical composition for causing hypertension.
8. 8. The kit of claim 7, further comprising a label or instructions for use of the kit in causing atrial fibrillation and hypertension in an animal subject.
9. 9. The kit of claim 8, wherein the label or instructions further prescribe use of the kit for preparing an animal model of ejection fraction retention heart failure.
10. 10. Use of an animal model of ejection fraction retention heart failure as claimed in claim 1 for mechanism research, medical device and drug development of ejection fraction retention heart failure diseases.

Description

Construction method and application of ejection fraction retention type heart failure animal model Technical Field The invention relates to the field of genetic engineering and experimental animal model design, in particular to an animal model for heart failure with reserved ejection fraction combining two phenotypes of atrial fibrillation and hypertension. Background Heart Failure (HF) is a complex clinical syndrome caused by impaired systole or blood filling. HF can be classified as either reduced ejection fraction (HFrEF), slightly reduced (HFmrEF) or reserved (HFpEF) HF, depending on the left ventricular contractile function. HFpEF accounts for 50% of all HF cases, with prevalence increasing with aging population. Over 4% of people 65 years old or older are affected by HFpEF with five year survival rates of about 50%, similar to HFrEF. HFpEF has a more complex etiology than HFrEF, and its symptoms are often manifested by left ventricular hypertrophy, increased left ventricular stiffness, and fluid retention. HFpEF is involved in a variety of risk factors and multiple organ complications, and the specific molecular pathogenic mechanisms are not well understood. The presently known drugs for treating HFrEF have no significant therapeutic effect on the mortality rate of HFpEF as a whole. Since clinical tissue samples of HFpEF patients are extremely limited, their mechanistic studies have been largely dependent on animal models. Currently, pressure overload caused by aortic systole (TAC) is the most widely used method of simulating ventricular hypertrophy and remodeling. However, the TAC model will inevitably develop into HFrEF, a phenomenon which rarely occurs in human HFpEF patients. Since HFpEF is often accompanied by various complications such as hypertension, diabetes, obesity, coronary artery disease, chronic kidney disease, and atrial fibrillation, combining hemodynamic and metabolic stress is a current common modeling strategy that includes a double-hit model of L-NAME induced Hypertension+high Fat Diet (HFD), a double-hit model of mild TAC+HFD, a triple-hit SAUNA model of saline-drinking water+unilateral nephrectomy+aldosterone, and a triple-hit model of HFD+deoxycorticosterone+aging (13 months). These methods provide valuable molecular insight into various pathological changes under different conditions, but these modeling strategies are generally long-lived (4-13 months after initiation of combination therapy), and the resulting pathological phenotype is not significant, often not prone to develop severe advanced heart failure and cause death. In view of the foregoing, there is a great need in the art for an animal model of heart failure with a short molding time that can produce a significant pathological phenotype with preserved ejection fraction. Disclosure of Invention The invention aims to provide a construction method and application of an animal model for heart failure with reserved ejection fraction. In a first aspect of the present invention, there is provided a method of constructing an animal model of ejection fraction retention heart failure, comprising the steps of: (a) Providing an animal subject, and (B) Atrial fibrillation and hypertension are caused to the individual animals. In another preferred embodiment, the animal subject is a non-human mammal, including bovine, ovine, canine, porcine, rabbit, non-human primate, rodent. In another preferred embodiment, the rodent comprises a monkey. In another preferred embodiment, the rodent comprises a mouse or a rat. In another preferred embodiment, the non-human mammal is a mouse. In another preferred embodiment, the method of causing atrial fibrillation comprises down-regulating Lkb gene expression in the atrium. In another preferred embodiment, the down-regulation comprises gene knockout or gene knockdown. In another preferred embodiment, the method of down-regulating Lkb gene expression is selected from the group consisting of: (A1) Introducing a deletion mutation or a frame shift mutation into Lkb gene; (A2) Carrying out gene editing on Lkb gene; (A3) RNAi interference is carried out on Lkb gene; (A4) Any combination of the A1 to A3. In another preferred embodiment, said down-regulating Lkb1 expression is performed by said gene editing knockout Lkb1 gene. In another preferred embodiment, the down-regulating Lkb1 expression in the atrium is performed by knocking out Lkb gene by a tissue-specific gene editing system. In another preferred embodiment, the tissue specific gene editing system is selected from the group consisting of: (B1) An atrial-specific promoter, a recombinase, a Lkb guide sequence expression cassette, and a reporter gene knockout system; (B2) An atrial-specific promoter, a ligand-inducible gene knockout system; or a combination thereof. In another preferred embodiment, the Lkb gene is knocked out using the tissue-specific gene editing system by a method selected from the group consisting of: (C1) Infecting an animal i