Methods for the treatment of heart failure with preserved ejection fraction

Abstract

The invention relates to the field of medicine, in particular of cardiology. The present invention is based on the identification of Runx 1 as a therapeutic target in heart failure with preserved ejection fraction (HFpEF)and addresses the pressing need for HFpEF therapies. Accordingly, the present invention relates to novel methods for the prophylaxis and treatment of HFpEF, and particularly, although not exclusively, to Runxf inhibitors for use in the prophylaxis and treatment of HFpEF. The invention provides methods of prophylaxis or treatment of HFpEF and compounds and compositions for use in those methods.

Description

[0001] METHODS FOR THE TREATMENT OF HEART FAILURE WITH PRESERVED EJECTION FRACTION Field of the Invention The invention relates to the field of medicine, in particular of cardiology. The invention is based on the identification of Runx1 as a therapeutic target in heart failure with preserved ejection fraction (HFpEF) and addresses the pressing need for HFpEF therapies. Accordingly, the present invention relates to novel methods for the treatment and prevention of HFpEF, and particularly, although not exclusively, to Runx1 inhibitors for use in the treatment and prevention of HFpEF. The invention provides methods of prophylaxis or treatment of HFpEF and compounds and compositions for use in those methods. Background Heart failure (HF), the leading cause of death world-wide, is a complex condition in which the heart is unable to meet the metabolic demands of the body. It is generally categorised by ejection fraction, which is the proportion of blood ejected from the left ventricle (LV) on each beat. Heart failure with reduced ejection fraction (HFrEF) involves a reduced capacity to eject blood from the heart. It is a common consequence of cardiac remodelling following myocardial infarction. HF with reduced ejection fraction (HFrEF) has been heavily investigated for many years and there are many treatment options available that reduce mortality (Tsao, C.W. et al. Temporal trends in the incidence of and mortality associated with heart failure with preserved and reduced ejection fraction. JACC Hear. Fail.6, 678~685 (2018)). More elusive, however, is HF with preserved ejection fraction (HFpEF) as it is increasing in prevalence, poorly understood, and very few treatment options are available as many of the classic HF treatments are not effective (Omote, K., Verbrugge, F.H. & Borlaug, B.A. Heart failure with preserved ejection fraction: mechanisms and treatment strategies. Annu. Rev. Med.73, 321~337 (2022)). In heart failure with preserved ejection fraction (HFpEF), the heart can pump but cannot lnklanhu nah]t, ok ep eoj^p ]^ha pk na-fill properly after contraction. HFpEF is typically not treatable with drugs that can be used for HFrEF. HFpEF is a multimorbidity syndrome, often developing alongside hypertension, metabolic stress, and diabetes, and includes both cardiac and systemic symptoms such as diastolic dysfunction, pulmonary oedema, hypertrophy, and exercise intolerance (Omote, K., Verbrugge, F.H. & Borlaug, B.A. Heart failure with preserved ejection fraction: mechanisms and treatment strategies. Annu. Rev. Med.73, 321~337 (2022)). There is significant pathophysiological heterogeneity in HFpEF, with a wide range of clinical presentations, and as such its pathophysiology is not clearly understood (Mishra, S. & Kass, D.A. Cellular and molecular pathobiology of heart failure with preserved ejection fraction. Nat. Rev. Cardiol.18, 400~423 (2021)). There is thus a desperate need for the development of new methods of treating HFpEF. Elucidating molecular or cellular factors contributing to the development of HFpEF is essential and is an important step toward identifying therapeutic targets. The present invention has been devised in light of the above considerations. Summary of the Invention The inventors have surprisingly found that reducing the expression or activity of Runx1 is effective in the prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF). Master-regulator transcription factor RUNX1 has been shown to be a key contributor to adverse cardiac remodelling following myocardial infarction (MI) and it may be a therapeutic target for treatment of adverse cardiac remodelling and progression to HF (McCarroll, C.S. et al. Runx1 deficiency protects against adverse cardiac remodeling after myocardial infarction. Circulation 137, 57~70 (2018); Riddell, A. et al. RUNX1: An emerging therapeutic target for cardiovascular disease. Cardiovasc. Res.116, 1410~1423 (2020)). Runx1 expression is consistently increased in key regions of remodelling and is implicated in pathology in HFrEF in animal models and humans. However, Runx1 is not consistently over-expressed in standard models of HFpEF in animal models. Until now, whether RUNX1 played any role in the pathophysiology of HFpEF was unknown. Despite the lack of overexpression in HFpEF disease models, the inventors have surprisingly found that Runx1 deficiency is protective in mice, and treatment by reducing the expression or activity of Runx1 (e.g. with shRNA (via adenovirus or AAV vectors), Ro5-3335 or Ro24-7429) delivers a therapeutic benefit in HFpEF (e.g. in improving diastolic function and reducing pulmonary oedema in particular). The invention therefore relates to the prophylaxis or treatment of HFpEF. More specifically, embodiments of the invention provide compounds, compositions and methods useful for the prophylaxis or treatment of HFpEF, based on reducing the expression or activity of Runx1. Accordingly, in a first aspect, the present invention provides a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), in a patient having or at risk of developing HFpEF, wherein the method comprises administering to the patient a therapeutically effective amount of an agent that decreases the expression or activity of Runx1 in the patient. The decrease in expression or activity of Runx1, may be a reduction in the expression or activity of Runx1 to any degree. In other words, Runx1 expression or activity may be around 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% of the level found in untreated cells or of the level found in a cell of a patient immediately prior to administration of the agent. Expression may be gene or protein expression and may be determined as described herein or by methods in the art that will be well known to a person skilled in the art. Suitable agents that decrease the expression or activity of Runx1 may be of any kind, including, for example, a therapeutic antibody, antibody fragment, antibody-like protein scaffold, aptamer, polypeptide, protein, genetic modifying agent, nucleic acid that specifically binds to a nucleic acid encoding Runx1, small interfering RNA (siRNA) configured to target Runx1, small hairpin RNA (shRNA) configured to target Runx1, a microRNA (miRNA) configured to target Runx1, an antisense oligonucleotide (ASO) configured to target Runx1, an RNA-guided endonuclease system configured to decrease the expression of Runx1, TALEN configured to target Runx1, a Runx1 inhibitor, small molecule inhibitor, small molecule degrader, or any combination thereof. In some embodiments, the agent that decreases the expression or activity of Runx1 in the patient may be a small molecule or an oligonucleotide. In some embodiments where the agent that decreases the expression or activity of Runx1 is an oligonucleotide (or nucleic acid), the agent may be selected from the group consisting of an agent that inhibits transcription of the gene encoding Runx1, an agent that inhibits post- transcriptional processing of RNA encoding Runx1, an agent that destabilises RNA encoding Runx1, and an agent that promotes the degradation of RNA encoding Runx1. In some embodiments, the agent is a siRNA, a shRNA, an miRNA, an antisense oligonucleotide (ASO), or an RNA-guided endonuclease system. Decreasing the expression or activity of Runx1 may comprise introducing a mutation or base edit that decreases Runx1 expression or effects a reduction or inhibition of the expression or activity of Runx1. Such a mutation or base edit may be introduced, for example, using a an RNA-guided endonuclease system, CRISPR-Cas system, an RNAi system, a TALEN, a Zn- finger nuclease, or a meganuclease. Such a mutation or base edit may be engineered using genome editing methods or by use of a CRISPR-Cas system, an inactivated CRISPR-Cas system, a Cas protein, a zinc finger protein (ZFP), a zinc finger nuclease (ZFN), a transcription activator-like effector (TALE), a transcription activator-like effector nuclease (TALEN), or a meganuclease. Such a mutation or base edit base edit may be made to genomic DNA or expressed RNA using a CRISPR-Cas system. In some embodiments, the agent is an ASO or an oligonucleotide which causes RNA interference (RNAi). The present invention contemplates the use of antisense nucleic acid to prevent, inhibit or reduce expression of Runx1. In some embodiments, an agent that decreases the expression of Runx1 in a patient may bring about a decrease in expression by RNA interference (RNAi). In some embodiments, the agent may be an inhibitory nucleic acid, such as antisense or small interfering RNA, including but not limited to shRNA or siRNA. In some embodiments, the inhibitory nucleic acid is provided in a vector. For example, in some embodiments the agent may be a Adeno-associated virus (AAV) vector encoding shRNA for Runx1 (i.e. configured to target human Runx1, e.g. human Runx1 X1 isoform (SEQ ID NO: 2)). More preferably the vector is an AAV9 viral vector. Oligonucleotide molecules, particularly RNA, may be employed to regulate gene expression. These include antisense oligonucleotides, targeted degradation of mRNAs by small interfering RNAs (siRNAs), post transcriptional gene silencing (PTGs), developmentally regulated sequence-specific translational repression of mRNA by micro-RNAs (miRNAs) and targeted transcriptional gene silencing. An antisense oligonucleotide (ASO) is an oligonucleotide, preferably single stranded that targets and binds, by complementary sequence binding, to a target oligonucleotide, e.g., mRNA. Where the target oligonucleotide is an mRNA, binding of the antisense to the mRNA blocks translation of the mRNA and expression of the gene product. Antisense oligonucleotides may be designed to bind sense genomic nucleic acid and inhibit transcription or promote degradation of a target nucleotide sequence. The nucleic acid sequences for Runx1 are known (exemplary mRNA sequence variants are available from GenBank® (for example under accession number XP_054180869 (SEQ ID NO: 2)), and so oligonucleotides may be designed to repress or silence the expression of Runx1. Such oligonucleotides may have any length, but may preferably be short, e.g., less than 100 nucleotides, e.g., 10-40 nucleotides, or 20-50 nucleotides, and may comprise a nucleotide sequence having complete- or near- complementarity (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% complementarity) to a sequence of nucleotides of corresponding length in the target oligonucleotide. The complementary region of the nucleotide sequence may have any length, but is preferably at least 5, and optionally no more than 50, nucleotides long, e.g. one of 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides. Where the target is human Runx1, the oligonucleotide preferably targets the X1 isoform (Genbank Accession: XP_054180869). Repression of Runx1 expression will preferably decrease the quantity of Runx1 expression by a cell, e.g., by a cardiac cell or cardiomyocyte cell. For example, in a given cell the repression of Runx1 by administration of a suitable nucleic acid will decrease the quantity of Runx1 expressed by the cell relative to an untreated cell of the same type (or relative to the same cell immediately prior to administration of the nucleic acid). Repression may be partial. Preferred degrees of repression are at least 50%, more preferably one of at least 60%, 70%, 80%, 85% or 90%. A level of repression between 90% and 100% is considered a ^silencing^ of expression or function. A role for the RNAi machinery and small RNAs in targeting of heterochromatin complexes and epigenetic gene silencing at specific chromosomal loci has been demonstrated. Double-stranded RNA (dsRNA)-dependent post transcriptional silencing, also known as RNA interference (RNAi), is a phenomenon in which dsRNA complexes can target specific genes of homology for silencing in a short period of time. It acts as a signal to promote degradation of mRNA with sequence identity. A 20-nt siRNA is generally long enough to induce gene-specific silencing, but short enough to evade host response. The decrease in expression of targeted gene products can be extensive with 90% silencing induced by a few molecules of siRNA. RNAi based therapeutics have been progressed into Phase I, II and III clinical trials for a number of indications (Nature 2009 Jan 22; 457(7228):426-433) and several RNAi based therapeutics are currently marketed as drugs approved by European and American regulatory agencies. In the art, these RNA sequences are termed ^short or small interfering RNAs^ (siRNAs) or ^microRNAs^ (miRNAs) depending on their origin. Both types of sequence may be used to down-regulate gene expression by binding to complementary RNAs and either triggering mRNA elimination (RNAi) or arresting mRNA translation into protein. siRNA are derived by processing of long double stranded RNAs and when found in nature are typically of exogenous origin. Micro-interfering RNAs (miRNA) are endogenously encoded small non- coding RNAs, derived by processing of short hairpins. Both siRNA and miRNA can inhibit the translation of mRNAs bearing partially complimentary target sequences without RNA cleavage and degrade mRNAs bearing fully complementary sequences. Accordingly, in some aspects the present invention provides the use of oligonucleotide sequences for downregulating the expression of Runx1 in order to treat HFpEF. siRNAs are typically double stranded and, to optimise the effectiveness of RNA mediated down-regulation of the function of a target gene, it is preferred that the length of the siRNA molecule is chosen to ensure correct recognition of the siRNA by the RISC complex that mediates the recognition by the siRNA of the mRNA target and so that the siRNA is short enough to reduce a host response. miRNAs are typically single stranded and have regions that are partially complementary enabling the ligands to form a hairpin. miRNAs are RNA genes which are transcribed from DNA but are not translated into protein. A DNA sequence that codes for a miRNA gene is longer than the miRNA. This DNA sequence includes the miRNA sequence and an approximate reverse complement. When this DNA sequence is transcribed into a single- stranded RNA molecule, the miRNA sequence and its reverse-complement base pair to form a partially double stranded RNA segment. The design of microRNA sequences is discussed in John et al, PLoS Biology, 11(2), 1862-1879, 2004. Typically, the siRNAs or miRNAs have between 10 and 40 ribonucleotides (or synthetic analogues thereof), more preferably between 17 and 30 ribonucleotides, more preferably between 19 and 25 ribonucleotides and most preferably between 21 and 23 ribonucleotides. In some embodiments of the invention employing double-stranded siRNA, the molecule may have symmetric 3' overhangs, e.g., of one or two (ribo)nucleotides, typically a UU of dTdT 3' overhang. Based on the disclosure provided herein, the skilled person can readily design suitable siRNA and miRNA sequences, for example using resources such the Ambion siRNA finder. siRNA and miRNA sequences can be synthetically produced and added exogenously to cause gene downregulation or produced using expression systems (e.g., vectors). In a preferred embodiment the siRNA is synthesized synthetically. Longer double stranded RNAs may be processed in the cell to produce siRNAs (see for example Myers (2003) Nature Biotechnology 21:324-328). The longer dsRNA molecule may have symmetric 3' or 5' overhangs, e.g., of one or two (ribo)nucleotides, or may have blunt ends. The longer dsRNA molecules may be 25 nucleotides or longer. Preferably, the longer dsRNA molecules are between 25 and 30 nucleotides long. More preferably, the longer dsRNA molecules are between 25 and 27 nucleotides long. Most preferably, the longer dsRNA molecules are 27 nucleotides in length. dsRNAs 30 nucleotides or more in length may be expressed using the vector pDECAP (Shinagawa et al., Genes and Dev., 17, 1340- 5, 2003). Another alternative is the expression of a short hairpin RNA molecule (shRNA) in the cell. shRNAs are more stable than synthetic siRNAs. A shRNA consists of short, inverted repeats separated by a small loop sequence. One inverted repeat is complimentary to the gene target. In the cell the shRNA is processed by DICER into a siRNA which degrades the target gene mRNA and suppresses expression. In a preferred embodiment, the shRNA is produced endogenously (within a cell) by transcription from a vector. shRNAs may be produced within a cell by transfecting the cell with a vector encoding the shRNA sequence under control of an RNA polymerase III promoter such as the human H1 or 7SK promoter or a RNA polymerase II promoter. As discussed elsewhere in this specification, it may be particularly advantageous to restrict the expression of the shRNA to cardiac cells (such as cardiomyocytes). One way of doing that is to transfect the cell with a vector encoding the shRNA sequence under control of a cardiac or cardiomyocyte specific promoter (examples of which are discussed elsewhere in this specification). Alternatively, the shRNA may be synthesised exogenously (in vitro) by transcription from a vector. The shRNA may then be introduced directly into the cell. Preferably, the shRNA molecule comprises a partial sequence of Runx1. Preferably, the shRNA sequence is between 40 and 100 bases in length, more preferably between 40 and 70 bases in length. The stem of the hairpin is preferably between 19 and 30 base pairs in length. The stem may contain G-U pairings to stabilise the hairpin structure. siRNA molecules, longer dsRNA molecules or miRNA molecules may be made recombinantly by transcription of a nucleic acid sequence, preferably contained within a vector. Preferably, the siRNA molecule, longer dsRNA molecule or miRNA molecule comprises a partial sequence of Runx1. In one embodiment, the siRNA, longer dsRNA or miRNA is produced endogenously (within a cell) by transcription from a vector. The vector may be introduced into the cell in any of the ways known in the art. Optionally, expression of the RNA sequence can be regulated using a tissue specific (e.g., cardiac specific) or cell specific (e.g. cardiomyocyte specific) promoter. In a further embodiment, the shRNA, siRNA, longer dsRNA, or miRNA is produced exogenously (in vitro) by transcription from a vector. Suitable vectors may be oligonucleotide vectors configured to express the oligonucleotide agent capable of Runx1 repression. Such vectors may be viral vectors or plasmid vectors. Preferably the vector is an AAV vector (e.g. AAV9) which is configured to express a shRNA targeting RunX1 (e.g. X1 isoform), most preferably in a cardiac specific or cardiomyocyte specific manner such that the dcrease in Runx1 expression occurs in a cardiac or cardiomyocyte specific manner. The therapeutic oligonucleotide may be incorporated in the genome of a viral vector and be operably linked to a regulatory sequence, e.g., promoter, sde_d `nerao epo atlnaooekj. Sda pani ^klan]^hu hejga`^ i]u ej_hq`a pda oepq]pekj sdana ] selected nucleotide sequence and regulatory nucleotide sequence are covalently linked in such a way as to place the expression of a nucleotide sequence under the influence or control of the regulatory sequence. Thus, a regulatory sequence is operably linked to a selected nucleotide sequence if the regulatory sequence is capable of effecting transcription of a nucleotide sequence which forms part or all of the selected nucleotide sequence. Viral vectors encoding promoter-expressed siRNA sequences are known in the art and have the benefit of long-term expression of the therapeutic oligonucleotide. Examples include lentiviral (Nature 2009 Jan 22; 457(7228):426-433), adenovirus (Shen et al., FEBS Lett 2003 Mar 27;539(1-3)111-4) and retroviruses (Barton and Medzhitov PNAS November 12, 2002 vol.99, no.2314943-14945). In other embodiments a vector may be configured to assist delivery of the therapeutic oligonucleotide to the site at which repression of Runx1 expression is required. Such vectors typically involve complexing the oligonucleotide with a positively charged vector (e.g., cationic cell penetrating peptides, cationic polymers and dendrimers, and cationic lipids); conjugating the oligonucleotide with small molecules (e.g., cholesterol, bile acids, sugars, and lipids), polymers, antibodies or antibody-like molecules, aptamers, and RNAs; or encapsulating the oligonucleotide in nanoparticulate formulations (Wang et al., AAPS J. 2010 Dec; 12(4): 492-503). In one embodiment, a vector may comprise a nucleic acid sequence in both the sense and antisense orientation, such that when expressed as RNA the sense and antisense sections will associate to form a double stranded RNA. Alternatively, siRNA molecules may be synthesized using standard solid or solution phase synthesis techniques which are known in the art. Linkages between nucleotides may be phosphodiester bonds or alternatives, for example, linking groups of the formula P(O)S, (thioate); P(S)S, (dithioate); P(O)NR'2; P(O)R'; P(O)OR6; CO; or CONR'2 wherein R is H (or a salt) or alkyl (1-12C) and R6 is alkyl (1-9C) is joined to adjacent nucleotides through-O-or- S-. Modified nucleotide bases can be used in addition to the naturally occurring bases and may confer advantageous properties on siRNA molecules containing them. For example, modified bases may increase the stability of the siRNA molecule, thereby reducing the amount required for silencing. The provision of modified bases may also provide siRNA ikha_qhao sde_d ]na ikna, kn haoo, op]^ha pd]j qjik`ebea` oeQM@. Sda pani ^ik`ebea` jq_hakpe`a ^]oa^ aj_kil]ooao jq_hakpe`ao sepd ] _kr]hajphu ik`ebea` ^]oa ]j` / kn oqc]r. For example, modified nucleotides include nucleotides having sugars which are covalently attached to low molecular weight organic groups other than a hydroxyl group at the 3'position and other than a phosphate group at the 5'position. Thus, modified nucleotides may also include 2'substituted sugars such as 2'-O-methyl; 2'-O-methoxyethyl; 2'-O-alkyl; 2'- O-allyl; 2'-S-alkyl; 2'-S-allyl; 2'-fluoro; 2'-halo or azido-ribose, carbocyclic sugar analogues, a- anomeric sugars; epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars, furanose sugars, and sedoheptulose. Modified nucleotides may also include phosphorodiamidate morpholino oligonucleotide (PMO). Modified nucleotides are known in the art and include alkylated purines and pyrimidines, acylated purines and pyrimidines, and other heterocycles. These classes of pyrimidines and purines are known in the art and include pseudoisocytosine, N4, N4-ethanocytosine, 8- hydroxy-N6-methyladenine, 4-acetylcytosine,5-(carboxyhydroxylmethyl) uracil, 5 fluorouracil, 5-bromouracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-carboxymethylaminomethyl uracil, dihydrouracil, inosine, N6-isopentyl-adenine, 1- methyladenine, 1-methylpseudouracil, 1-methylguanine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3- methylcytosine, 5-methylcytosine, N6-methyladenine, 7-methylguanine, 5- methylaminomethyl uracil, 5-methoxy amino methyl-2-thiouracil, -D-mannosylqueosine, 5- methoxycarbonylmethyluracil, 5methoxyuracil, 2 methylthio-N6-isopentenyladenine, uracil-5- oxyacetic acid methyl ester, psueouracil, 2-thiocytosine, 5-methyl-2 thiouracil, 2-thiouracil, 4- thiouracil, 5methyluracil, N-uracil-5-oxyacetic acid methylester, uracil 5-oxyacetic acid, queosine, 2-thiocytosine, 5-propyluracil, 5-propylcytosine, 5-ethyluracil, 5ethylcytosine, 5- butyluracil, 5-pentyluracil, 5-pentylcytosine, and 2,6,diaminopurine, methylpsuedouracil, 1- methylguanine, 1-methylcytosine. Methods relating to the use of RNAi to silence genes in C. elegans, Drosophila, plants, and mammals are known in the art (Fire A, et al., 1998 Nature 391:806-811; Fire, A. Trends Genet.15, 358-363 (1999); Sharp, P. A. RNA interference 2001. Genes Dev.15, 485-490 (2001); Hammond, S. M., et al., Nature Rev. Genet.2, 110-1119 (2001); Tuschl, T. Chem. Biochem.2, 239-245 (2001); Hamilton, A. et al., Science 286, 950-952 (1999); Hammond, S. M., et al., Nature 404, 293-296 (2000); Zamore, P. D., et al., Cell 101, 25-33 (2000); Bernstein, E., et al., Nature 409, 363-366 (2001); Elbashir, S. M., et al., Genes Dev.15, 188- 200 (2001); WO0129058; WO9932619, and Elbashir S M, et al., 2001 Nature 411:494-498). Accordingly, the invention provides a nucleic acid that is capable, when suitably introduced into or expressed within a mammalian, e.g., human, cell that otherwise expresses Runx1, of suppressing Runx1 expression by RNAi. The nucleic acid may have substantial sequence identity to a portion of DNA, pre-mRNA or mRNA encoding for Runx1. The nucleic acid may be a double-stranded siRNA. As the skilled person will appreciate, ]j` ]o atlh]eja` bqnpdan ^ahks, ] oeQM@ ikha_qha i]u ]hok ej_hq`a ] odknp 3^ CM@ sequence. Alternatively, the nucleic acid may be a DNA (usually double-stranded DNA) which, when transcribed in a mammalian cell, yields an RNA having two complementary portions joined via a spacer, such that the RNA takes the form of a hairpin when the complementary portions hybridise with each other. In a mammalian cell, the hairpin structure may be cleaved from the molecule by the enzyme DICER, to yield two distinct, but hybridised, RNA molecules. It is expected that perfect identity / complementarity between the nucleic acid of the invention and the target sequence, although preferred, is not essential. Accordingly, the nucleic acid of the invention may include a single mismatch compared to the mRNA of Runx1. It is expected, however, that the presence of even a single mismatch is likely to lead to reduced abbe_eaj_u, ok pda ]^oaj_a kb ieoi]p_dao eo lnabanna`. Vdaj lnaoajp, 3^ krand]jco i]u ^a excluded from the consideration of the number of mismatches. Sda pani ^_kilhaiajp]nepu^ eo jkp heiepa` pk _kjrajpekj]h ^]oa l]enejc ^apsaaj jq_hae_ ]_e` consisting of naturally occurring ribo- and / or deoxyribonucleotides, but also includes base pairing between mRNA and nucleic acids of the invention that include non-natural nucleotides. The strands that form the double-opn]j`a` QM@ i]u d]ra odknp 3^ `ejq_hakpe`a krand]jco, sde_d i]u ^a CM@ kn QM@. Sda qoa kb ] 3^ CM@ krand]jc d]o jk abba_p kj oeQM@ ]_perepu _kil]na` pk ] 3^ QM@ krand]jc ^qp na`q_ao pda _kop kb _daie_]h oujpdaoeo kb pda jq_hae_ acid strands (Elbashir et al., 2001c). For this reason, DNA dinucleotides may be preferred. When present, the dinucleotide overhangs may be symmetrical to each other, though this is jkp aooajpe]h. Hj`aa`, pda 3^ krand]jc kb pda oajoa (qllan) opn]j` eo ennahar]jp bkn QM@e activity, as it does not participate in mRNA recognition and degradation (Elbashir et al., 2001a, 2001b, 2001c). Vdeha QM@e atlaneiajpo ej Cnkokldeh] odks pd]p ]jpeoajoa 3^ krand]jco i]u l]npe_el]pa ej iQM@ na_kcjepekj ]j` p]ncapejc (Dh^]oden ap ]h. 2001_), 3^ krand]jco `k jkp ]lla]n pk ^a necessary for RNAi activity of siRNA in mammalian cells. Incorrect annealing kb 3^ overhangs is therefore thought to have little effect in mammalian cells (Elbashir et al.2001c; Czauderna et al.2003). Any dinucleotide overhang may therefore be used in the antisense strand of the siRNA. Nevertheless, the dinucleotide is preferably -UU or ~UG (or ~TT or ~TG if the overhang is DNA), more preferably -UU (or ~TT). The ~UU (or ~TT) dinucleotide overhang is most effective and is consistent with (i.e., capable of forming part of) the RNA polymerase III end of transcription signal (the terminator signal is TTTTT). Accordingly, this dinucleotide is most preferred. The dinucleotides AA, CC and GG may also be used, but are less effective and _kjoamqajphu haoo lnabanna`. Lknakran, pda 3^ krand]jco i]u ^a kieppa` ajpenahu bnki pda siRNA. Disclosed herein are single-stranded nucleic acids (herein referred to as single-stranded siRNAs) respectively consisting of a component strand of one of the aforementioned double- opn]j`a` jq_hae_ ]_e`o, lnaban]^hu sepd pda 3^-overhangs, but optionally without. Also described are kits containing pairs of such single-stranded nucleic acids, which are capable of hybridising with each other in vitro to form the aforementioned double-stranded siRNAs, which may then be introduced into cells. Disclosed herein is DNA that, when transcribed in a mammalian cell, yields an RNA (herein also referred to as an shRNA) having two complementary portions which are capable of self- hybridising to produce a double-stranded motif, or a sequence that differs from any one of the aforementioned sequences by a single base pair substitution. The complementary portions will generally be joined by a spacer, which has suitable length and sequence to allow the two complementary portions to hybridise with each other. The psk _kilhaiajp]nu (e.a., oajoa ]j` ]jpeoajoa) lknpekjo i]u ^a fkeja` 5^-3^ ej either order. The spacer will typically be a short sequence, of approximately 4-12 nucleotides, preferably 4-9 nucleotides, more preferably 6-9 jq_hakpe`ao. Onaban]^hu pda 5^ aj` kb pda ol]_an (eiia`e]pahu 3^ kb pda qlopna]i _kilhaiajp]nu lknpekj) _kjoeopo kb pda jq_hakpe`ao ~UU- or ~UG-, again preferably ~UU- (though, again, the use of these particular dinucleotides is not essential). A suitable spacer, recommended for use in the pSuper system of OligoEngine (Seattle, Washington, USA) is UUCAAGAGA. In this and other cases, the ends of the spacer may hybridise with each other by a small number (e.g., 1 or 2) of base pairs. Reieh]nhu, pda pn]jo_ne^a` QM@ lnaban]^hu ej_hq`ao ] 3^ krand]jc bnki pda `ksjopna]i complementary portion. Again, this is preferably ~UU or ~UG, more preferably ~UU. Such shRNA molecules may then be cleaved in the mammalian cell by the enzyme DICER to yield a double-stranded siRNA as described above, in which one or each strand of the du^ne`eoa` `oQM@ ej_hq`ao ] 3^ krand]jc. Techniques for the synthesis of the nucleic acids of the invention are of course well known in the art. The skilled person is well able to construct suitable transcription vectors for the DNA of the invention using well-known techniques and commercially available materials. In particular, the DNA will be associated with control sequences, including a promoter and a transcription termination sequence. Various suitable systems are available. For example, the pSuper and pSuperior systems of OligoEngine (Seattle, Washington, USA). These use a polymerase-III promoter (H1) and a S5 pn]jo_nelpekj paniej]pkn oamqaj_a pd]p _kjpne^qpao psk T naoe`qao ]p pda 3^ aj` kb pda transcript (which, after DICER processing, lnkre`a ] 3^ TT krand]jc kb kja opn]j` kb pda siRNA). Another suitable system is described in Shin et al. (RNA, 2009 May; 15(5): 898- 910), which uses another polymerase-III promoter (U6). The double-stranded siRNAs of the invention may be introduced into mammalian cells in vitro or in vivo using known techniques, as described below, to suppress expression of Runx1. Similarly, transcription vectors containing the DNAs of the invention may be introduced into cells in vitro or in vivo using known techniques, as described below, for transient or stable expression of RNA, again to suppress expression of Runx1. Accordingly, the invention also provides a method of suppressing Runx1 expression in a mammalian, e.g., human, cell, the method comprising administering to the cell a double- stranded siRNA of the invention or a transcription vector of the invention. Similarly, the invention further provides a method of treating the diseases described herein, the method comprising administering to a subject a double-stranded siRNA of the invention or a transcription vector of the invention. Materials and methods suitable for the administration of siRNA duplexes and DNA vectors of the invention are well known in the art and improved methods are under development, given the potential of RNAi technology. Generally, many techniques are available for introducing nucleic acids into mammalian cells. The choice of technique will depend on whether the nucleic acid is transferred into cultured cells in vitro or in vivo in the cells of a patient. Techniques suitable for the transfer of nucleic acid into mammalian cells in vitro include the use of liposomes, electroporation, microinjection, cell fusion, DEAE-dextran and calcium phosphate precipitation. In vivo gene transfer techniques include transfection with viral (typically retroviral) vectors, exosomes or exosome-like particles, naked oligonucleotides (e.g. linked to targeting molecules such as GalNAc (N-acetylgalactosamine) or pegylated oligonucleotides), lipid nanoparticles, and viral coat protein-liposome mediated transfection (Dzau et al. (2003) Trends in Biotechnology 11, 205-210; Roberts TC et al., Nature Rev. Drug Discov., Vol 19, 2020; Paunovska K, Nature Rev. Genetics, Vol 23, 2022). In particular, suitable techniques for cellular administration of the nucleic acids of the invention both in vitro and in vivo are disclosed in the following articles: General reviews: Borkhardt, A.2002. Blocking oncogenes in malignant cells by RNA interference--new hope for a highly specific cancer treatment? Cancer Cell.2:167-8. Hannon, G.J.2002. RNA interference. Nature.418:244-51. McManus, M.T., and P.A. Sharp. 2002. Gene silencing in mammals by small interfering RNAs. Nat Rev Genet.3:737-47. Scherr, M., M.A. Morgan, and M. Eder.2003b. Gene silencing mediated by small interfering RNAs in mammalian cells. Curr Med Chem.10:245-56. Shuey, D.J., D.E. McCallus, and T. Giordano.2002. RNAi: gene-silencing in therapeutic intervention. Drug Discov Today. 7:1040-6. Systemic delivery using liposomes: Lewis, D.L., J.E. Hagstrom, A.G. Loomis, J.A. Wolff, and H. Herweijer.2002. Efficient delivery of siRNA for inhibition of gene expression in postnatal mice. Nat Genet.32:107-8. Paul, C.P., P.D. Good, I. Winer, and D.R. Engelke.2002. Effective expression of small interfering RNA in human cells. Nat Biotechnol.20:505-8. Song, E., S.K. Lee, J. Wang, N. Ince, N. Ouyang, J. Min, J. Chen, P. Shankar, and J. Lieberman.2003. RNA interference targeting Fas protects mice from fulminant hepatitis. Nat Med.9:347-51. Sorensen, D.R., M. Leirdal, and M. Sioud.2003. Gene silencing by systemic delivery of synthetic siRNAs in adult mice. J Mol Biol.327:761-6. Virus mediated transfer: Abbas-Terki, T., W. Blanco-Bose, N. Deglon, W. Pralong, and P. Aebischer.2002. Lentiviral-mediated RNA interference. Hum Gene Ther.13:2197-201. Barton, G.M., and R. Medzhitov.2002. Retroviral delivery of small interfering RNA into primary cells. Proc Natl Acad Sci U S A.99:14943-5. Devroe, E., and P.A. Silver.2002. Retrovirus-delivered siRNA. BMC Biotechnol.2:15. Lori, F., P. Guallini, L. Galluzzi, and J. Lisziewicz.2002. Gene therapy approaches to HIV infection. Am J Pharmacogenomics. 2:245-52. Matta, H., B. Hozayev, R. Tomar, P. Chugh, and P.M. Chaudhary.2003. Use of lentiviral vectors for delivery of small interfering RNA. Cancer Biol Ther.2:206-10. Qin, X.F., D.S. An, I.S. Chen, and D. Baltimore.2003. Inhibiting HIV-1 infection in human T cells by lentiviral-mediated delivery of small interfering RNA against CCR5. Proc Natl Acad Sci U S A.100:183-8. Scherr, M., K. Battmer, A. Ganser, and M. Eder.2003a. Modulation of gene expression by lentiviral-mediated delivery of small interfering RNA. Cell Cycle.2:251-7. Shen, C., A.K. Buck, X. Liu, M. Winkler, and S.N. Reske.2003. Gene silencing by adenovirus-delivered siRNA. FEBS Lett.539:111-4. Peptide delivery: Morris, M.C., L. Chaloin, F. Heitz, and G. Divita.2000. Translocating peptides and proteins and their use for gene delivery. Curr Opin Biotechnol.11:461-6. Simeoni, F., M.C. Morris, F. Heitz, and G. Divita.2003. Insight into the mechanism of the peptide-based gene delivery system MPG: implications for delivery of siRNA into mammalian cells. Nucleic Acids Res.31:2717-24. Other technologies that may be suitable for delivery of siRNA to the target cells are based on nanoparticles or nanocapsules such as those described in US patent numbers 6,649,192B and 5,843,509B. The agent may be an RNA-guided endonuclease system. In some embodiments, the agent is an RNA-guided endonuclease system comprising (a) an RNA-guided endonuclease; and (b) a guide RNA complementary to a target sequence within the target gene (i.e., Runx1) in the genome of the target cell. In some embodiments, the agent is an RNA-guided endonuclease system comprising (a) a nucleic acid sequence encoding an RNA-guided endonuclease; and (b) a nucleic acid sequence encoding a guide RNA complementary to a target sequence within the target gene (i.e., Runx1) in the genome of the target cell. Sda lnaoajp ejrajpekj i]u ]hok qoa ] BQHROQ (^_hqopana` nacqh]nhu ejpanol]_a` odknp l]hej`nkie_ nala]po^) ouopai pk `a_na]oa atlnaooekj kb pda p]ncap cajao. Sda BQHROQ kn CRISPR-Cas system is derived from a prokaryotic RNA-guided defence system. There are at least eleven different CRISPR-Cas systems, which have been grouped into three major types (I-III). Type II and type III CRISPR-Cas systems have been adapted as a genome- engineering tool. Typically, most naturally occurring type II CRISPR-Cas systems employ three components: ^ a protein endonuclease Cas (CRISPR-associated protein) having DNA nickase activity which is referred to in this specification as an RNA-guided endonuclease (or an RNA-guided DNA endonuclease), ^ ] ^p]ncapejc^ kn ^cqe`a^ QM@ (BQHROQ-RNA or crRNA) comprising a short sequence, typically of approximately 20nucleotides, complementary to a target sequence (^lnkpkol]_an^) ej pda cajkia, ]j` ^ ] ^o_]bbkh`^ QM@ (pn]jo-acting CRISPR RNA or tracrRNA) which interacts with the crRNA and recruits the Cas endonuclease. In some embodiments, the system introduces a DNA break into the target gene. The target gene may be knocked-out (i.e., made inoperative). In other embodiments of the invention, the endonuclease is non-functional and is used to target the DNA or RNA to reduce expression by means other than introducing DNA cuts and is instead functionalized to inhibit expression of encoded proteins by e.g., inhibition of translation or transcription. Typically, assembly of these components and hybridisation of the crRNA with its target sequence in the chromosome results in cleavage of the chromosome by the endonuclease, at or close to the target sequence. Cleavage also requires that the target DNA contains a recognition site for the Cas enzyme (protospacer adjacent motif, or PAM) located sufficiently close to the crRNA target sequence, typically immediately adjacent the 3^ aj` kb pda p]ncap sequence. Cellular repair of the DNA break can lead to the insertion / deletion / mutation of bases and mutation at the target locus. This three-component system has been simplified by fusing together crRNA and tracrRNA, to create a chimeric single guide RNA (sgRNA or gRNA). Hybridisation of the gRNA with the target sequence leads to cleavage of the target DNA at an adjacent / upstream PAM site. An gRNA can therefore be regarded as comprising a crRNA component (which determines the target sequence) and a tracrRNA component (which recruits the endonuclease). The protein component of the CRISPR system is referred to as an endonuclease and may have enzymatic activity (i.e., DNA nickase activity) when associated with the appropriate RNA factors. Typically, the endonuclease will cleave chromosomal DNA. In some embodiments, the endonuclease is a Cas9 protein. Examples include Staphylococcus aureus (SaCas9), Streptococcus pyogenes (SpCas9), Neisseria meningitidis (NM Cas9), Streptococcus thermophilus (ST Cas9), Treponema denticola (TD Cas9), or variants thereof. The PAM sequences recognised by these enzymes are well known in the art. Beneficially, SaCas9, CjCas9, and NmCas9 (2.9~3.3 kb) each allow for the packaging of both Cas9 and gRNA in a single AAV vector. When using a catalytically active endonuclease, the target sequence recognised by the guide RNA may be upstream of a suitable site for insertion. The endonuclease may comprise a nuclear localisation sequence (NLS) effective in mammalian cells, such as the SV40 large T antigen NLS, which has the sequence PKKKRKV. Other mammalian NLS sequences are known to the skilled person. The endonuclease may comprise multiple copies of an NLS, e.g., two or three copies of an NLS. Where multiple NLS sequences are present, they are typically repeats of the same NLS. In some embodiments, a gene encoding the endonuclease component of the system will be under transcriptional control of an RNA polymerase II promoter e.g., a viral or human RNA polymerase II promoter. Examples include CMV or SV40 promoter, or a mammalian ^dkqoagaalejc^ lnkikpan. Fajao aj_k`ejc ]ju QM@ _kilkjajpo (cQM@, _nQM@ kn tracrRNA) will typically be under the transcriptional control of an RNA polymerase III promoter (e.g., a human RNA polymerase UII promoter) such as the U6 or H1 promoter, or variants thereof which retain or have enhanced activity. In some embodiments, the agent is a CRISPR-Cas vector system. In some embodiments, the agent is a CRISPR-Cas vector system comprising (a) a Cas endonuclease; and (b) a guide RNA complementary to a target sequence within the target gene (i.e., Runx1). In some embodiments, the agent is a CRISPR-Cas vector system comprising (a) a nucleic acid sequence encoding a Cas endonuclease; and (b) a nucleic acid sequence encoding a guide RNA complementary to a target sequence within the target gene (i.e., Runx1). Hj okia ai^k`eiajpo, pda atlnaooekj harah kb Qqjt1 i]u ^a na`q_a` ^u ] ^gjk_g`ksj^ approach. In some embodiments, the expression level of Runx1 may be knocked out entirely. That may be particularly desirable in scenarios where the expression of Runx1 is attenuated in a cell-specific manner, e.g. a cardiomycete-specific manner. Knockdown or knockout of RUNX1 can be determined by many methods known in the art. For example, by quantitative immunoblotting (per Janes K.A. An analysis of critical factors for quantitative immunoblotting. Sci. Signal.2015; 8 (rs2)). For example, with the following antibodies: RUNX1 (Cell Signaling #4336, 1:1000), RUNX2 (MBL International #D130-3, 1:1000), BAE^ (R]jp] Bnqv Aekpa_djkhkceao $RB-20693, 1:1000), vinculin (Millipore #05- 386, 1:10,000), tubulin (Abcam #ab89984, 1:20,000), and GAPDH (Ambion #AM4300, 1:20,000). Alternatively, or in addition, knockdown of RUNX1 can be determined by qrtPCR or RNAseq data or RNAscope. In some embodiments, Runx1 has an amino acid sequence of SEQ ID NO: 1. In some embodiments, Runx1 has a transcript sequence of SEQ ID NO: 2. In some embodiments, the activity or expression of Runx1 is decreased by at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or by 100% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50- 60%, 60-70%, 70-80%, 80-90% or 90-100% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50-90%, 50-80%, 50-70% or 50-60% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50-100%, 60-90% or 70-80% compared to a baseline or control value. Throughout this specification, a ^nalnaooekj^, ^reduction^ or ^decrease^ in the level of the activity or expression of Runx1 may refer to either a reduction compared to a baseline value (e.g. a starting value prior to administration of the agent) or a reduction compared to a control (such as, for example, placebo, a value for an untreated cell of the same type, or an average value for a patient population suffering from HFpEF). For example, administration of a Runx1 inhibitor or a construct comprising a nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 may reduce Runx1 levels by decreasing Runx1 levels below a previously determined baseline level (for example, a starting value established prior to administration of the Runx1 inhibitor or construct). Alternatively, administration of a Runx1 inhibitor or a construct comprising a nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 may reduce Runx1 levels by causing a greater decrease than a placebo at a specific timepoint after administration. Appropriate control values may include for example, those of an untreated control (or an untreated control population), those resulting from administration of a placebo, an average value associated with a patient with HFpEF or of a population of patients with HFpEF. Alternatively the corresponding value of an untreated cell of the same type may be a useful control value, for example, in situations such as those where the nucleic acid is expressed in a cell-specific manner. In the case of a Runx1 inhibitor, the most convenient control comparison may be that of a placebo. In the case of a shRNA vector, an appropriate control comparison may be e.g. the same vector featuring a non-targeted sequence, for example a ^o_n]i^ha^ oamqaj_a e.g. AAV9-Runx1-shRNA vs control = AAV9-scramble-shRNA). In some embodiments, the decrease in activity or expression of Runx1 is confined to cardiomyocytes. In some embodiments, the agent is a Runx1 inhibitor. For example, the Runx1 inhibitor may be a:

[0002] , wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof. In certain embodiments this formula can be interpreted as:

[0003] T y be: wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3). The Runx1 inhibitor may be:

[0004] wherein X is N(R,R1) or O R and R1are each independently C1-C4alkyl or wherein one of R and R1is hydrogen, preferably wherein N(R,R1) is NH(CH3). The Runx1 inhibitor may be: The Runx1 inhibitor may be Ro5-3335 or Ro24-7429, or a combination thereof. The Runx1 inhibitor may be administered intravenously, subcutaneously or orally. Preferably, the Runx1 inhibitor is administered orally. The Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor may be administered as a single dose or in multiple doses. In some embodiments, the method comprises administration of the Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor at a dosage of 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day or 400 mg / day. Preferably, the treatment is administered regularly, preferably between several times a day and every week, more preferably every day or every other day. In a preferred embodiment, the treatment is administered once a day. The duration of treatment with a Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor typically lasts as long as the HFpEF persists. In some embodiments, the method comprises administration of the Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor daily or every second day until symptoms resolve. In some embodiments, the treatment may be a lifelong treatment. In some embodiments, the duration of treatment with a Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor is optionally between 1 day and 52 weeks, preferably between 1 day and 25 weeks. The amount of Runx1 inhibitor to be administered has to be determined by standard procedure well known to those of ordinary skill in the art. Physiological data of the patient (e.g. age, size, weight, and physical general condition) and the routes of administration have to be taken into account to determine the appropriate dosage, so a therapeutically effective amount will be administered to the patient. The Runx1 inhibitor may be administered alone, or in combination with one or more other active ingredients, including e.g. other Runx1 inhibitors or other cardiac medicines. The Runx1 inhibitor may be administered in a pharmaceutical composition. The pharmaceutical compositions that comprise the Runx1 inhibitor may be formulated in various dosage forms for oral or parenteral administration. The pharmaceutical compositions may comprise a Runx1 inhibitor as an active ingredient, alone or in combination with a pharmaceutically acceptable vehicle, carrier, diluent, or excipient, or a mixture thereof. The pharmaceutical compositions may also be formulated as modified release dosage forms, including delayed-, extended-, prolonged-, sustained-, pulsatile-, controlled-, accelerated- and fast-, targeted-, or programmed-release dosage forms. These dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art (see e.g. Remington: The Science and Practice of Pharmacy, supra; Modified-Release Drug Deliver Technology, Rathbone et al., Eds., Drugs and the Pharmaceutical Science, Marcel Dekker, Inc.: New York, N.Y., 2003; Vol.126). Preferably, the pharmaceutical compositions that comprise the Runx1 inhibitor comprise at least one excipient or pharmaceutically acceptable carrier. Conventional excipients can be used according to techniques well known by those skilled in the art. The pharmaceutical compositions comprising a Rux1 inhibitor can be formulated in accordance with standard pharmaceutical practice (see e.g. Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York). In preferred embodiments, the pharmaceutical compositions comprising a Runx1 inhibitor are provided in a dosage form for oral administration, which comprise the Runx1 inhibitor, and one or more pharmaceutically acceptable excipients or carriers. The pharmaceutical compositions comprising a Runx1 inhibitor provided herein may be provided in solid, semisolid, or liquid dosage forms for oral administration. As used herein, oral administration also includes buccal, lingual, and sublingual administration. Suitable oral dosage forms include, but are not limited to, tablets, capsules, pills, troches, lozenges, pastilles, cachets, pellets, medicated chewing gum, granules, bulk powders, effervescent or non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers, sprinkles, elixirs, and syrups. In addition to the active ingredient(s), the pharmaceutical compositions may contain one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, binders, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, colouring agents, dye-migration inhibitors, sweetening agents, and flavouring agents. In other embodiments, the pharmaceutical compositions comprising a Runx1 inhibitor are provided in a dosage form for parenteral administration (e.g. for injection or infusion), which comprise the Runx1 inhibitor, and one or more pharmaceutically acceptable excipients or carriers. The pharmaceutical compositions comprising a Runx1 inhibitor may be administered parenterally by injection, infusion, or implantation, for local or systemic administration. Parenteral administration, as used herein, include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial, intravesical, and subcutaneous administration. The pharmaceutical compositions comprising a Runx1 inhibitor may be formulated in any dosage forms that are suitable for parenteral administration, including solutions, suspensions, emulsions, micelles, liposomes, microspheres, nanosystems, and solid forms suitable for solutions or suspensions in liquid prior to injection. Such dosage forms can be prepared according to conventional methods known to those skilled in the art of pharmaceutical science (see, Remington: The Science and Practice of Pharmacy, supra). In another aspect, the invention provides a composition comprising an agent that decreases the expression or activity of Runx1 for use in a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), in a patient having or at risk of developing HFpEF, wherein the method comprises administering a therapeutically effective amount of the composition to the subject. In some embodiments, Runx1 has an amino acid sequence of SEQ ID NO: 1 (i.e. human Runx1 amino acid sequence). In some embodiments, the activity or expression of Runx1 is decreased by at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or by 100% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50- 60%, 60-70%, 70-80%, 80-90% or 90-100% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50-90%, 50-80%, 50-70% or 50-60% compared to a baseline or control value. Optionally, the activity or expression of Runx1 is decreased in the range 50-100%, 60-90% or 70-80% compared to a baseline or control value. Throughout this specification, a ^nalnaooekj^, ^reduction^ or ^decrease^ in the level of the activity or expression of Runx1 may refer to either a reduction compared to a baseline value (e.g. a starting value prior to administration of the agent) or a reduction compared to a control (such as e.g. placebo placebo or or an average value for a patient population suffering from HFpEF). For example, administration of a Runx1 inhibitor or a construct comprising a nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 may reduce Runx1 levels by decreasing Runx1 levels below a previously determined baseline level (for example, a starting value established prior to administration of the Runx1 inhibitor or construct). Alternatively, administration of a Runx1 inhibitor or a construct comprising a nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 may reduce Runx1 levels by causing a greater decrease than a placebo at a specific timepoint after administration. Appropriate control values may include for example, those of an untreated control (or an untreated control population), those resulting from administration of a placebo, an average value associated with a patient with HFpEF or of a population of patients with HFpEF. For example, in the case of a Runx1 inhibitor, an appropriate control comparison may be a placebo. In the case of a shRNA vector, an appropriate control comparison may be the o]ia ra_pkn ba]pqnejc ] ^o_n]i^ha^ oamqaj_a e.g. AAV9-Runx1-shRNA vs control = AAV9- scramble-shRNA). In some embodiments, the agent that decreases the expression or activity of Runx1 is a Runx1 inhibitor. ^Hjde^epkn^ ]o qoa` danaej nabano pk ]ju _kilkqj` kn lnk`nqc pdanakb pd]p inhibits or blocks the activity of Runx1. The inhibition can be direct or indirect. In preferred embodiments, the agent that decreases the expression or activity of Runx1 is a Runx1 inhibitor and the inhibitor decreases the expression or activity of Runx1 compared with a placebo and / or compared with the expression or activity of Runx1 immediately prior to administration of the inhibitor. In more preferred embodiments, the inhibitor decreases the expression or activity of Runx1 compared with a placebo. In particularly preferred embodiments, the inhibitor decreases the activity of Runx1. In preferred embodiments, the Runx1 inhibitor is a small molecule inhibitor. For example, the Runx1 inhibitor may be:

[0005] , wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof. In certain embodiments this formula can be interpreted as:

[0006] T y be: wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3). The Runx1 inhibitor may be:

[0007] wherein X is N(R,R1) or O R and R1are each independently C1-C4alkyl or wherein one of R and R1is hydrogen, preferably wherein N(R,R1) is NH(CH3) In preferred embodiments, the Runx1 inhibitor may be: In some embodiments, the Runx1 inhibitor is Ro5-3335 or Ro24-7429, or a combination thereof. In some embodiments, the method comprises intravenous, subcutaneous or oral administration of the Runx1 inhibitor. Preferably, the method comprises oral administration of the Runx1 inhibitor. More preferably, the Runx1 inhibitor is administered orally. Preferably, the method comprises administration of the Runx1 inhibitor as a single dose or in multiple doses. In some embodiments, the method comprises administration of the Runx1 inhibitor at a dosage of 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day or 400 mg / day. Preferably, the Runx1 inhibitor is administered regularly, preferably between several times a day and every week, more preferably every day or every other day. In a preferred embodiment, the Runx1 inhibitor is administered once a day. The duration of treatment with a Runx1 inhibitor or the pharmaceutical composition comprising a Runx1 inhibitor typically lasts as long as the HFpEF persists. In some embodiments, the method comprises administration of the Runx1 inhibitor daily or every second day until symptoms resolve. In some embodiments, the treatment may be a lifelong treatment. In some embodiments, the duration of treatment with a Runx1 inhibitor is optionally between 1 day and 52 weeks, preferably between 1 day and 25 weeks. The amount of Runx1 inhibitor to be administered has to be determined by standard procedure well known to those of ordinary skill in the art. Physiological data of the patient (e.g. age, size, weight, and physical general condition) and the routes of administration have to be taken into account to determine the appropriate dosage, so a therapeutically effective amount will be administered to the patient. In some embodiments, the agent that decreases the expression or activity of Runx1 is a nucleic acid that is configured to target human Runx1, e.g. human Runx1 X1 isoform (e.g. SEQ ID NO: 2)). In some embodiments, the agent that decreases the expression or activity of Runx1 is a nucleic acid that is configured to reduce or repress the expression of human Runx1 (e.g. human Runx1 X1 isoform (e.g. SEQ ID NO: 2)) compared to a baseline value or control value. Preferably, the agent is a small hairpin RNA (shRNA), a small interfering RNA (siRNA), a microRNA (miRNA), an antisense oligonucleotide (ASO), or an RNA-guided endonuclease system configured to decrease the expression of human Runx1 (e.g. human Runx1 X1 isoform (SEQ ID NO: 2)) compared to a baseline value or control value. More preferably, the agent is a shRNA configured to reduce or repress the expression of human Runx1 (e.g. human Runx1 X1 isoform (e.g. SEQ ID NO: 2)) compared to a baseline value or control value. In some embodiments, the agent that decreases the expression or activity of Runx1 is a nucleic acid that specifically binds to a nucleic acid encoding Runx1. Such an agent, may, for example, specifically bind to an mRNA transcript encoding Runx1 or a gene encoding Runx1 (e.g. human Runx1 X1 isoform (SEQ ID NO: 2)). Preferably, the agent is capable of binding specifically to a human mRNA transcript encoding Runx1. More preferably, the agent that decreases the expression or activity of Runx1 specifically binds to a nucleic acid encoding Runx1 having a sequence of SEQ ID NO: 2 (Runx1 (Isoform X1) AML1c human transcript sequence). Whilst in some situations it may be useful to generally decrease the level or activity of Runx1 in the patient, it may also be helpful to control the location, timing and / or amount of any decrease in activity or expression. For example, in some embodiments, the decrease in activity or expression of Runx1 may be usefully confined to cells of the heart, such as cardiomyocytes. That may be helpful, for example, in reducing or avoiding unwanted side effects. With that in mind, in some cases, the agent that decreases the expression or activity of Runx1 may be specifically localised to cardiomyocytes. For example, in embodiments where the agent that decreases the expression or activity of Runx1 is a nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1, the expression of the agent may be restricted to cardiac cells or cardiomyocytes. Cardiac- specific or cardiomyocyte-specific expression can be achieved by various means. Typically, the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is under the control of a cardiac or cardiomyocyte specific promoter. For example, the promoter controlling the expression of the nucleic acid agent may be cardiomyocyte specific, such that the agent is expressed only in cardiomyocytes. Suitable promoters may be constitutive, whereby they direct expression under most environmental conditions or developmental stages, developmental stage specific or inducible. The use of inducible or regulatable promoters adds another layer of control over the timing and / or amount of transgene expression and can further increase the safety of the construct. Where the vector used to deliver the transgene is an AAV vector and cardiac specific expression of the transgene for knocking down expression of Runx1 is desired, several convenient promoters for AAV are available (for example, those discussed in Pacak & Byrne. AAV vectors for cardiac gene transfer: experimental tools and clinical opportunities. Mol Ther. 2011 Sep;19(9):1582-90. doi: 10.1038 / mt.2011.124. Epub 2011 Jul 26.). Other suitable promoters may include HIF regulated promoters for hypoxic response (e.g. MLC, MHC, MyoD, MEF2). Preferably, the promoter is inducible, initiating transcription in cardiomyocytes in response to environmental, chemical or developmental stimuli. Suitable promoters may include but are not limited to Tet-Off and Tet-On. The Tet-Off and Tet-On systems allow modulation of a gene-of-interest by administration or withdrawal of tetracyclines. The Tet-Off system allows silencing of gene expression by administration of tetracycline or tetracycline-derivatives like doxycycline, whereas the Tet-On system allows activation of gene expression by doxycycline. In some embodiments, the nucleic acid that specifically binds to a nucleic acid encoding Runx1 is a small hairpin RNA (shRNA), small interfering RNA (siRNA) or morpholino oligonucleotide. Optionally, the nucleic acid that specifically binds to a nucleic acid encoding Runx1 is configured to target a sequence according to SEQ ID NO: 2 (or a corresponding sequence). The nucleic acid agents that specifically bind to a nucleic acid encoding Runx1 as described herein, and / or a regulatory sequence are preferably provided as part of an expression cassette to enable expression of the nucleic acid agent in a patient. Suitable expression cassettes for use in the present invention may be constructed by standard techniques known in the art, to comprise 5' and 3' regulatory sequences, including, but not limited to promoter sequences, terminator fragments, polyadenylation sequences or enhancer sequences operably linked to the sequences of interest. Such elements may be included in the expression construct to obtain the optimal expression and function of the nucleic acid agent, in the patient (and therefore to control the expression of Runx1 in the patient in the desired manner). In addition, polynucleotides encoding, for example, selectable markers and reporter genes may be included. Also provided in the expression cassette may be transcription and translation initiation regions, to enable expression of the incoming genes, transcription and translational termination regions, and regulatory sequences. These sequences may be native to the species of patient (for example, human sequences), or may be heterologous. The expression cassettes may be a bi-functional expression cassette which functions in multiple hosts (for example to facilitate construction and testing of the cassette as well as optimal expression in the patient). A regulatory sequence is a nucleotide sequence which is capable of influencing transcription or translation of a gene or gene product, for example in terms of initiation, accuracy, rate, stability, downstream processing and mobility. Examples of regulatory sequences include promoters, 5' and 3' UTR's, enhancers, transcription factor or protein binding sequences, start sites and termination sequences, ribosome binding sites, recombination sites, polyadenylation sequences, sense or antisense sequences. The regulatory sequences may be derived from the same species as the patient or may be derived from another organism as required. In some embodiments, the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is provided in a vector. The vector may be chosen in order to optimise expression in the patient. Preferably, the vector is a viral vector. More preferably the vector is an Adeno-associated virus (AAV) vector. Even more preferably the vector is an AAV9 viral vector. The compositions for use in a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF) preferably comprise one or more pharmaceutically acceptable carriers, excipients or diluents. Preferably, the compositions are pharmaceutical compositions. In a further aspect, the present invention provides a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), wherein the method comprises administering to a patient having or at risk of developing HFpEF a therapeutically effective amount of a compound according to the formula:

[0008] , wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof. In certain embodiments this formula can be interpreted as:

[0009] In some embodiments, the compound may be a compound according to the formula: wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3). In some embodiments, the compound may be a compound according to the formula: wherein X is N(R,R1) or O R and R1are each independently C1-C4alkyl or wherein one of R and R1is hydrogen, preferably wherein N(R,R1) is NH(CH3). In some embodiments, the compound may be a compound according to the formula: In some embodiments, the compound may is Ro5-3335 or Ro24-7429, or a combination thereof. Preferably, the method comprises administration of the compound as a single dose or in multiple doses. In some embodiments, the method comprises intravenous, subcutaneous or oral administration of the compound. Preferably, the method comprises oral administration of the compound. More preferably, the compound is administered orally. In some embodiments, the method comprises administration of the compound at a dosage of 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day or 400 mg / day. Preferably, the compound is administered regularly, preferably between several times a day and every week, more preferably every day or every other day. In a preferred embodiment, the compound is administered once a day. The duration of treatment with the compound typically lasts as long as the HFpEF persists. In some embodiments, the compound is administered daily or every second day until symptoms resolve. In some embodiments, the treatment may be a lifelong treatment. In some embodiments, the duration of treatment is optionally between 1 day and 52 weeks, preferably between 1 day and 25 weeks. The amount of compound to be administered has to be determined by standard procedure well known to those of ordinary skill in the art. Physiological data of the patient (e.g. age, size, weight, and physical general condition) and the routes of administration have to be taken into account to determine the appropriate dosage, so a therapeutically effective amount will be administered to the patient. In preferred embodiments, the compound is administered as part of a pharmaceutical composition. In a further aspect, the present invention provides a compound for use in a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), wherein the method comprises administering to a patient having or at risk of developing HFpEF a therapeutically effective amount of the compound, wherein the compound is a compound according to the formula: , wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof. In certain embodiments this formula can be interpreted as: In some embodiments, the compound is a compound according to the formula: wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N-(R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N-(R2,R3). In some embodiments, the compound is a compound according to the formula: wherein X is N(R,R1) or O R and R1are each independently C1-C4alkyl or wherein one of R and R1is hydrogen, preferably wherein N(R,R1) is NH(CH3). In some embodiments, the compound is a compound according to the formula: In some embodiments, the compound is Ro5-3335 or Ro24-7429, or a combination thereof. The compound may be administered intravenously, subcutaneously or orally. Preferably, the compound is administered orally. The compound may be administered as a single dose or in multiple doses. In some embodiments, the method comprises administration of the compound at a dosage of 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day or 400 mg / day. Preferably, the compound is administered regularly, preferably between several times a day and every week, more preferably every day or every other day. In a preferred embodiment, the compound is administered once a day. The duration of treatment with a compound typically lasts as long as the HFpEF persists. In some embodiments, the method comprises administration of the compound daily or every second day until symptoms resolve. In some embodiments, the treatment may be a lifelong treatment. In some embodiments, the duration of treatment with a compound is optionally between 1 day and 52 weeks, preferably between 1 day and 25 weeks. The amount of compound to be administered has to be determined by standard procedure well known to those of ordinary skill in the art. Physiological data of the patient (e.g. age, size, weight, and physical general condition) and the routes of administration have to be taken into account to determine the appropriate dosage, so a therapeutically effective amount will be administered to the patient. The compound may be administered alone, or in combination with one or more other active ingredients, including e.g. other cardiac medicines. The compound may be administered in a pharmaceutical composition. In preferred embodiments, the compound is administered as part of a pharmaceutical composition. The pharmaceutical compositions that comprise the compound may be formulated in various dosage forms for oral or parenteral administration. The pharmaceutical compositions may comprise the compound as an active ingredient, alone or in combination with a pharmaceutically acceptable vehicle, carrier, diluent, or excipient, or a mixture thereof. The pharmaceutical compositions may also be formulated as modified release dosage forms, including delayed-, extended-, prolonged-, sustained-, pulsatile-, controlled-, accelerated- and fast-, targeted-, or programmed-release dosage forms. These dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art (see e.g. Remington: The Science and Practice of Pharmacy, supra; Modified-Release Drug Deliver Technology, Rathbone et al., Eds., Drugs and the Pharmaceutical Science, Marcel Dekker, Inc.: New York, N.Y., 2003; Vol. 126). Preferably, the pharmaceutical compositions that comprise the compound comprise at least one excipient or pharmaceutically acceptable carrier. Conventional excipients can be used according to techniques well known by those skilled in the art. The pharmaceutical compositions comprising the compound can be formulated in accordance with standard pharmaceutical practice (see e.g. Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York). In preferred embodiments, the pharmaceutical compositions comprising the compound are provided in a dosage form for oral administration, which comprise the comprise, and one or more pharmaceutically acceptable excipients or carriers. The pharmaceutical compositions comprising the compound provided herein may be provided in solid, semisolid, or liquid dosage forms for oral administration. As used herein, oral administration also includes buccal, lingual, and sublingual administration. Suitable oral dosage forms include, but are not limited to, tablets, capsules, pills, troches, lozenges, pastilles, cachets, pellets, medicated chewing gum, granules, bulk powders, effervescent or non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers, sprinkles, elixirs, and syrups. In addition to the active ingredient(s), the pharmaceutical compositions may contain one or more pharmaceutically acceptable carriers or excipients, including, but not limited to, binders, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, colouring agents, dye-migration inhibitors, sweetening agents, and flavouring agents. In other embodiments, the pharmaceutical compositions comprising the compound are provided in a dosage form for parenteral administration (e.g. for injection or infusion), which comprise the compound, and one or more pharmaceutically acceptable excipients or carriers. The pharmaceutical compositions comprising the compound may be administered parenterally by injection, infusion, or implantation, for local or systemic administration. Parenteral administration, as used herein, include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial, intravesical, and subcutaneous administration. The pharmaceutical compositions comprising the compound may be formulated in any dosage forms that are suitable for parenteral administration, including solutions, suspensions, emulsions, micelles, liposomes, microspheres, nanosystems, and solid forms suitable for solutions or suspensions in liquid prior to injection. Such dosage forms can be prepared according to conventional methods known to those skilled in the art of pharmaceutical science (see, Remington: The Science and Practice of Pharmacy, supra). Medical Uses With the knowledge of the unexpectedly key role of Runx1 in the pathogenesis of HFpEF and with data demonstrating prophylaxis and treatment of HfpEF in a mouse model, the agents, compounds, or compositions as described herein that reduce Runx1 signalling are expected to be efficacious for the treatment of HFpEF. Accordingly, the invention provides the agents, compounds, or compositions described herein for use in methods of treatment or prophylaxis of HFpEF. The invention further provides methods of treatment or prophylaxis of HFpEF comprising administering one or more of the agents, compounds, or compositions as described herein to a patient. The invention further provides the use of an agent, compound, or composition as described herein for the manufacture of a medicament for the treatment or prophylaxis of HFpEF in a patient. HFpEF Patients Heart failure can occur in the presence of a normal or a reduced left ventricular ejection fraction. ^Ejection Fraction^ refers to the division of the left ventricle stroke volume by its end-diastolic volume. In other words, the term ^ejection fraction^ refers to the percentage of blood that is pumped out of a filled ventricle with each heartbeat. Methods for measuring the ejection fraction, for example with an echocardiogram, are well known to a person skilled in the art. As used herein, the terms ^Heart Failure with Preserved Ejection Fraction^ (GElDE), ^heart failure with normal ejection fraction^ or ^diastolic heart failure (DHF)^, are equivalent and refer to a patient that suffers from heart failure and that does not present a significant reduction of their left ventricular ejection fraction compared to that of a control (e.g., a healthy individual) or compared to an average value from a healthy population. In certain embodiments, an ejection fraction is classified as normal and a patient suffering from HF is classified as having a HFpEF when the ejection fraction is superior or equal to 40%, preferably superior or equal to 45%, more preferably superior or equal to 50%, still more preferably superior or equal to 55%, yet more preferably superior or equal to 60%, even more preferably 70%. HFpEF usually occurs in older patients with risk factors such as obesity, diabetes and hypertension and is more common in women. The methods according to the invention (and compounds and compositions for use in those methods) are aimed to treat a patient affected with HFpEF or at risk of developing HFpEF. In all aspects of the invention (i.e. all methods, compounds, compositions described herein), the patient to be treated may be an animal, preferably a mammal, even more preferably a human. As used herein, the terms ^subject^, ^individual^ or ^patient^ are interchangeable and refer to an animal, preferably to a mammal, even more preferably to a human. The human patient according to the invention may be a child, an infant, an adolescent or an adult, preferably an adult. Preferably, the patient is a woman. However, the patient can also be a non-human animal, in particular mammals such as dogs, cats, horses, cows, pigs, sheep and non-human primates, among others, that are in need of treatment or at risk of developing HFpEF. In preferred embodiments the patient is an adult human patient having HFpEF. The human patient according to the invention is preferably an adult of at least 40 years old, more preferably an adult of at least 50 years old, still more preferably an adult of at least 60 years old, yet more preferably an adult of at least 65 years old, even more preferably an adult of at least 70 years old. In a most preferred embodiment, the human patient according to the invention is an adult of at least 75 years old. The patient may be currently diagnosed with HFpEF or exhibiting one or more conditions associated with HFpEF, a patient who has been diagnosed with or exhibited one or more conditions associated with HFpEF in the past, or a patient who has been deemed at risk of developing HFpEF in the future. The patient may be suffering from HFpEF or may have been identified as being at risk of developing HFpEF. Preferably, the patient has been diagnosed with HFpEF. Typically, a patient is considered as being affected with HFpEF when they suffer from heart failure and present a preserved ejection fraction, in particular they do not present a significant reduction of their left ventricular ejection fraction compared to that of a control (e.g., a healthy individual) or compared to an average value from a healthy population. Preferably, the ejection fraction of the patient suffering from HFpEF is superior or equal to about 40%, preferably superior or equal to about 45%, more preferably superior or equal to about 50%. Alternatively, the ejection fraction of the patient suffering from HFpEF is superior or equal to about 55%), preferably superior or equal to about 60%. Accordingly, prior to administration of the compounds or compositions disclosed herein the left ventricular ejection fraction (LVEF) of the patient is superior or equal to 40%, optionally to 50%, preferably to 55%, preferably to 60%, even more preferably to 65%, most preferably to 70%. In some situations a patient with HFpEF may present an elevated B-type natriuretic peptide (BNP) level in combination with a normal ejection fraction. Preferably, a patient has an elevated BNP level when they present a BNP blood concentration superior to about 35 pg / ml, preferably superior to about 50 pg / ml, more preferably superior to about 75 pg / ml, still more preferably superior to about 100 pg / ml, even more preferably superior to about 200 pg / ml. A patient may be considered as being affected with HFpEF when they present evidence of cardiac dysfunction, such as abnormal left ventricular filling and / or elevated filling pressures. Accordingly, prior to administration of the compounds or compositions disclosed herein the patient may display evidence of cardiac dysfunction characteristic of HFpEF, such as abnormal left ventricular filling and / or elevated filling pressures. Echocardiography techniques can be used to diagnose diastolic dysfunction and require measurement of multiple echocardiographic parameters to confirm an HFpEF diagnosis. So, a combined approach is needed. Multiple echocardiographic parameters have been proposed as sensitive and specific, including mitral inflow velocity patterns, pulmonary vein flow patterns, E:A reversal, tissue Doppler measurements (i.e., E / E' ratio), and M-mode echo measurements (i.e. of left atrial size). Algorithms have further been developed which combine multiple echocardiographic parameters. A person skilled in the art is able to choose the most appropriate parameters and / or algorithms. There are four basic echocardiographic patterns of diastolic dysfunction, which are graded from I to IV: - pda ieh`aop bkni eo _]hha` ]j ^]^jkni]h nah]t]pekj l]ppanj^, kn cn]`a H `e]opkhe_ `uobqj_pekj. On the mitral inflow Doppler echocardiogram, there is reversal of the normal E / A ratio. This pattern may develop normally with age in some patients, and many grade I patients will not have any clinical signs or symptoms of heart failure; grade II diastolic dysfunction is called ^loaq`k jkni]h behhejc `uj]ie_o^. Sdeo eo _kjoe`ana` ik`an]pa `e]opkhe_ `uobqj_pekj ]j` eo associated with elevated left atrial filling pressures. These patients more commonly have symptoms of heart failure, and many have left atrial enlargement due to the elevated pressures in the left heart; cn]`ao HHH ]j` HU `e]opkhe_ `uobqj_pekj ]na _]hha` ^naopne_pera behhejc `uj]ie_o^. These are both severe forms of diastolic dysfunction, and patients tend to have advanced heart failure symptoms: x cn]`a HHH `e]opkhe_ `uobqj_pekj l]peajpo sehh `aikjopn]pa narano]h kb pdaen `e]opkhe_ abnormalities on echocardiogram when they perform the Valsalva manoeuvre. This is nabanna` pk ]o ^naranoe^ha naopne_pera `e]opkhe_ `uobqj_pekj^; x cn]`a HU `e]opkhe_ `uobqj_pekj l]peajpo sehh jkp `aikjopn]pa naranoe^ehepu kb pdaen echocardiogram abnormalities, and are therefore said to suffer from ^beta`, naopne_pera `e]opkhe_ `uobqj_pekj^. The presence of either grade III or IV diastolic dysfunction is associated with a poor prognosis. These patients will have left atrial enlargement. Accordingly, in a particular embodiment, a patient may be considered as being affected with HFpEF when they present with a heart failure and a diastolic dysfunction, preferably of at least grade I, more preferably of at least grade II, even more preferably of at least grade III. In a most preferred embodiment, a patient is considered as being affected with HFpEF when they present with a heart failure and a diastolic dysfunction of grade IV. In some embodiments, prior to administration of the agent, compound or composition the E / A ratio of the patient is >2.5, optionally >3.0, optionally >3.5. In some embodiments, prior to administration of the agent, compound or composition the patient has an End Diastolic Pressure Volume Relationship (EDPVR) of >12 mmHg, optionally 12-18 iiGc, lnaban]^hu |18 iiGc. It will be appreciated that diagnosis of HFpEF can be complex and in certain situations it may be helpful to use a combination of the measures and thresholds disclosed herein. A person skilled in the art is able to choose the most appropriate parameters in the circumstances. It may also be helpful to use an integrated measure of various symptoms. One such measure is the HFA-PEFF score. Another such measure is the H2FPEF score. Accordingly, in some embodiments, prior to administration of the compound or composition the patient has an H2EODE o_kna kb |6 kn ]j GE@-ODEE o_kna kb |5, lnaban]^hu sdanaej pda patient has an H2FPEF score of 6-9 or an HFA-PEFF score of 5-6. In certain situations, invasive exercise testing may be required to determine whether a patient has HFpEF. In some embodiments, HFpEF may be diagnosed by cardiac catheterization showing increased LV filling pressures. Accordingly, prior to administration of the compound or composition the patient may have elevated pulmonary capillary wedge pressure (PCWP) at rest or during exercise. In some embodiments, prior to administration of the compound or composition the patient may have a rest pulmonary capillary wedge lnaooqna (OBVO) |15 ii Gc kn atan_eoa OBVO |25 ii Gc. Treatment or Prophylaxis of HFpEF The inventors have surprisingly discovered that decreasing the level of Runx1 expression or activity results in an improvement in diastolic function. Accordingly, the methods according to the invention (and agents, compounds and compositions for use in those methods) are aimed to treat a patient affected with HFpEF or at risk of developing HFpEF. Accordingly, the administration of an agent, compound or composition disclosed herein in accordance with those methods may result in an improvement in one or more symptoms of HFpEF in a patient to values not characteristic of HFpEF. For example, the method may result in an improvement in, restoration of, or prevention of deterioration in either diastolic function or cardiac compliance. The administration of an agent, compound or composition disclosed herein may result in an improvement in diastolic function or cardiac compliance compared to a baseline value or a control value. Preferably, the improvement in diastolic function or cardiac compliance is an improvement compared with a placebo. The administration of an agent, compound or composition disclosed herein may result in a complete restoration of diastolic function or cardiac compliance to values not characteristic of HFpEF. In cases where the patient is suffering from HFpEF, the administration of an agent, compound or composition disclosed herein may result in an improvement in diastolic function or cardiac compliance compared with baseline. In cases where the patient is at risk of developing HFpEF the administration of an agent, compound or composition disclosed herein may result in a preservation of diastolic function in the normal range of values for a healthy patient or may reduces the deterioration of diastolic function associated with HFpEF compared with a control, baseline or reference value. Diastolic function can be determined by many methods known in the art. Established measures of diastolic function include E to A wave ratio, for example, from M-mode echocardiography or end-diastolic pressure-volume relationship (EDPVR). The E / A ratio is a marker of the function of the left ventricle of the heart. It represents the ratio of peak velocity blood flow from left ventricular relaxation in early diastole (the E wave) to peak velocity flow in late diastole caused by atrial contraction (the A wave). Diastolic function or cardiac compliance (i.e. how easily a chamber of the heart or the lumen of a blood vessel expands when it is filled with a volume of blood) may also be measured by End Diastolic Pressure Volume Relationship (EDPVR), for example, the shkla (^) kb pda aj`- diastolic pressure volume relationship. In some embodiments, the administration of an agent, compound or composition disclosed herein results in a restoration of diastolic function or cardiac compliance to values not characteristic of HFpEF, preferably wherein diastolic function or cardiac compliance is measured by E to A wave ratio from M-mode echocardiography and / or End Diastolic Pressure Volume Relationship (EDPVR). In some embodiments, the administration of an agent, compound or composition disclosed herein results in an E / A ratio of 0.5-2.5, preferably 0.75~2.0, more preferably 0.75~1.5. In some embodiments, the administration of an agent, compound or composition disclosed herein results in an improvement in End Diastolic Pressure Volume Relationship (EDPVR) to a value not characteristic of HFpEF. Preferably, the administration of an agent, compound or composition disclosed herein results in an End Diastolic Pressure Volume Relationship (EDPVR) of <12 mmHg. More preferably, the administration of an agent, compound or composition disclosed herein results in an End Diastolic Pressure Volume Relationship (EDPVR) of <10 mmHg. Even more preferably, the administration of an agent, compound or composition disclosed herein results in an End Diastolic Pressure Volume Relationship (EDPVR) of 3~8 mmHg. In some embodiments, the administration of an agent, compound or composition disclosed herein results in an improvement in pulmonary oedema, resolution of pulmonary oedema or inhibits the development of pulmonary oedema. In some embodiments, the administration of an agent, compound or composition disclosed herein results in an improvement in H2FPEF score and / or HFA-PEFF score. Preferably, the administration of an agent, compound or composition disclosed herein results in an improvement in H2FPEF score and / or HFA-PEFF score to a value not characteristic of HFpEF. Preferably, the administration of an agent, compound or composition disclosed herein results in an H2FPEF score of <6 and / or an HFA-PEFF score of <5. More preferably, the administration of an agent, compound or composition disclosed herein results in an H2FPEF score of <5 or an HFA-PEFF score of <4. Even more preferably, the administration of an agent, compound or composition disclosed herein results in an H2FPEF score of <4 or an HFA-PEFF score of <3. In some embodiments, the administration of an agent, compound or composition disclosed herein results in an improvement in rest pulmonary capillary wedge pressure (PCWP) or exercise PCWP. That is, a reduction in rest pulmonary capillary wedge pressure (PCWP) or exercise PCWP. Preferably, the administration of an agent, compound or composition disclosed herein results in an improvement in rest pulmonary capillary wedge pressure (PCWP) and / or exercise PCWP score to a value not characteristic of HFpEF. More preferably, the administration of an agent, compound or composition disclosed herein results in a rest PCWP <15 mm Hg and / or exercise PCWP <25 mm Hg. Even more preferably, the administration of an agent, compound or composition disclosed herein results in a rest PCWP <12 mm Hg and / or exercise PCWP <25 mm Hg. For the avoidance of doubt, as used herein, the term ^treatment^, ^treat" or ^treating^ refers to any act intended to ameliorate the health status of patients such as therapy, retardation, delaying the worsening of a disease or disorder. The terms "treat," "treating," or "treatment" as used herein with regard to a condition refers to preventing the condition, slowing the onset or rate of development of the condition, reducing the risk of developing the condition, preventing or delaying the development of symptoms associated with the condition, reducing or ending symptoms associated with the condition, generating a complete or partial regression of the condition, lessening the severity of the condition, changing the levels of one or more markers associated with the condition, or some combination thereof. In certain embodiments, such term refers to the amelioration or eradication of HFpEF or symptoms associated with HFpEF. In other embodiments, this term refers to minimizing the worsening of HFpEF resulting from the administration of one or more therapeutic agents to a subject with HFpEF. For example, with regard to HFpEF, "treatment" may refer e.g. to a improvement in diastolic function, cardiac compliance or exercise tolerance or reduction in pulmonary oedema, or some combination thereof. The term ^lnklduh]teo^ refers to inhibiting the onset of HFpEF and / or its attendant symptoms or barring a subject from developing HFpEF. ^Onklduh]teo^ i]u ej_hq`a reducing the probability of developing or redeveloping HFpEF, or of a recurrence of previously-controlled HFpEF, in a subject who does not have, but is at risk of or is susceptible to, redeveloping HFpEF or a recurrence of the condition. ^Onklduh]teo^ may also include delaying the onset of HFpEF and / or its attendant symptoms and reducing a l]peajp^s risk of developing HFpEF. Throughout, a ^pdan]laqpe_]hhu abba_pera ]ikqjp^ kb ] compound or composition as used in the methods according to the invention is an amount of a compound or composition that produces a desired therapeutic effect in a patient, such as e.g. treating one or more of the symptoms of HFpEF. The precise therapeutically effective amount is an amount of the a compound or composition that will yield the most effective results in terms of therapeutic efficacy in a given patient. This amount will vary depending upon a variety of factors, including but not limited to the characteristics of the therapeutic composition (including, e.g., activity, pharmacokinetics, pharmacodynamics, and bioavailability), the physiological condition of the subject (including, e.g., age, body weight, sex, disease type and stage, medical history, general physical condition, responsiveness to a given dosage, and other present medications), the nature of the pharmaceutically acceptable carrier or carriers in the composition, and the route of administration. One skilled in the clinical and pharmacological arts will be able to determine a therapeutically effective amount through routine experimentation, namely by monitoring a l]peajp^s response to administration of a composition and adjusting the dosage accordingly. For additional guidance, see, e.g., Remington: The Science and Practice of Pharmacy, 21st Edition, Univ. of Sciences in Philadelphia (USIP), Lippincott Williams & Wilkins, Philadelphia, PA, 2005, and Goodman & Gilman's The Pharmacological Basis of Therapeutics, 11th Edition, McGraw-Hill, New York, NY, 2006. The invention includes the combination of the aspects and preferred features described except where such a combination is clearly impermissible or expressly avoided. Brief Description of the Figures Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures in which: Figure 1 shows Runx1-deficient mice are protected against HFpEF phenotype. Panel a) shows a schematic of two-hit protocol and experimental groups. Panel b) shows Body weight over the experimental protocol in each of the groups. * p < 0.05 for 2HM-Runx1fl / flcompared to CTRL- Runx1fl / fl, # p < 0.05 for 2HM-Runx1^ / ^compared to CTRL-Runx1^ / ^, ]j` w l < 0.05 bkn 2GL-Runx1^ / ^compared to 2HM-Runx1fl / flby mixed-effects analysis. Panel c) shows systolic blood pressure (BP) over the experimental protocol. * p < 0.05 for 2HM-Runx1fl / flcompared to CTRL-Runx1fl / fland # p < 0.05 for 2HM-Runx1^ / ^compared to CTRL-Runx1^ / ^by mixed-effects analysis. Panels d)- i) show characterisation of the HFpEF phenotype by d) exercise intolerance testing quantified by running distance, e) pulmonary oedema quantified by wet to dry lung weight ratio, hypertrophy quantified by f) left ventricular (LV) weight normalised to tibial length (TL) g) cardiomyocyte cross sectional area and diastolic function quantified by h) E to A wave ratio from M-mode echocardiography and i) pda ohkla (^) kb pda aj`-diastolic pressure volume relationship. Figure 2 shows AAV9-mediated knockdown of Runx1 protects against diastolic dysfunction. Panel a) shows a schematic of two-hit protocol and experimental groups. Panel b) shows Body weight over the experimental protocol i* p < 0.05 for HFD+L-NAME compared to NFD+Water, # p < 0.05 for AAV9-scramble-shRNA (HFD+L-NAME) compared to MEC+V]pan, ]j` w l < 0.05 bkn @@U9-Runx1-shRNA (HFD+L-NAME) compared to NFD+Water by mixed-effects analysis. Panel c) shows systolic blood pressure (BP) over the experimental protocol. * p < 0.05 for HFD+L-NAME compared to NFD+Water, # p < 0.05 for AAV9-scramble-shRNA (HFD+L-M@LD) _kil]na` pk MEC+V]pan, ]j` w l < 0.05 bkn @@U9- Runx1-shRNA (HFD+L-NAME) compared to NFD+Water by mixed-effects analysis. Panels d)- j) show characterisation of the HFpEF phenotype by d) exercise intolerance testing quantified by running distance, e) pulmonary oedema quantified by wet to dry lung weight ratio, hypertrophy quantified by f) left ventricular (LV) weight normalised to tibial length (TL), g) and diastolic function quantified by E to A wave ratio from M mode echocardiography h) kj pda bej]h saag ]j` e) kran pda atlaneiajp]h lnkpk_kh ]j` f) pda ohkla (^) kb pda aj`- diastolic pressure volume relationship. Figure 3 shows Runx1 small molecule inhibitor Ro5-3335 remedies against diastolic dysfunction. Panel a) shows a schematic of two-hit protocol and experimental groups. Panels b)-f) show characterisation of the HFpEF phenotype by b) exercise intolerance testing quantified by running distance, c) pulmonary oedema quantified by wet to dry lung weight ratio, d) hypertrophy quantified by left ventricular (LV) weight normalised to tibial length (TL); and diastolic function quantified by e) E to A wave ratio from M mode a_dk_]n`ekcn]ldu ]j` b) pda ohkla (^) kb pda aj`-diastolic pressure volume relationship. Figure 4 shows Runx1 small molecule inhibitor Ro24-7429 remedies against diastolic dysfunction. Panel a) shows a schematic of two-hit protocol and experimental groups. Panels b)-e) show characterisation of the HFpEF phenotype by b) exercise intolerance testing quantified by running distance, c) pulmonary oedema quantified by wet to dry lung weight ratio, d) hypertrophy quantified by left ventricular (LV) weight normalised to tibial length (TL), and e) diastolic function quantified by E to A wave ratio from M mode echocardiography. Figure 5 shows a map of pZacf-U6-luc-ZsGreen used to produce recombinant AAV vectors. Polynucleotide and Amino Acid Sequences Below are exemplary polynucleotide and amino acid sequences used in accordance with the invention. Table 1. sequences and accession numbers used in the current study (see NCBI- GenBank Flat File Release 256.0), updated June 152023). SEQ ID NO: ACCESSION DESCRIPTION 1 XP_054180869 Human Runx1 (Isoform X1) AML1c amino acid sequence 2 XP_054180869 Human Runx1 (Isoform X1) AML1c transcript sequence 3 NM_001111023 Amino Acid Sequence of Mouse Runx1 (AML1b) 4 NM_001111023 Transcript sequence of Mouse Runx1 (AML1b) 5 NP_001104491 Amino acid sequence of Mouse Runx1 (Longest isoform (AML1c) Transcript variant 1. This variant has different start site and different N-terminus compared to AML1b). 6 NM_001111021 Mus musculus runt related transcription factor 1 (Runx1), transcript variant 1, mRNA. 7-Runx1 targeting sequence8 - shRNA sequence used in examples. (Hairpin Loop Sequence: CTCGAG. and TTTTT terminates the shRNA expression for U6 promoter) 9 - Scramble control shRNA sequence used in examples. 10 - Runx1 shRNA short hairpin forward primer sequence 11 - Runx1 shRNA short hairpin reverse primer sequence 12 - Scramble shRNA short hairpin forward primer sequence 13 - Scramble shRNA short hairpin reverse primer sequence 14 - pZacf-U6-luc-ZsGreen used to produced recombinant AAV vectors - full sequence (plasmid map shown in Figure 4) SEQ ID NO: 1 Human Runx1 (Isoform X1) AML1c LOCUS XP_054180869 480 aa linear PRI 20-MAR-2023 DEFINITION runt-related transcription factor 1 isoform X1 [Homo sapiens]. ACCESSION XP_054180869 VERSION XP_054180869.1 DBLINK BioProject: PRJNA807723 DBSOURCE REFSEQ: accession XM_054324894.1 KEYWORDS RefSeq. SOURCE Homo sapiens (human) Human Runx1 (Isoform X1) AML1c amino acid sequence: 1 masdsifesf psypqcfmre cilgmnpsrd vhdastsrrf tppstalspg kmsealplga 61 pdagaalagk lrsgdrsmve vladhpgelv rtdspnflcs vlpthwrcnk tlpiafkvva 121 lgdvpdgtlv tvmagndeny saelrnataa mknqvarfnd lrfvgrsgrg ksftltitvf 181 tnppqvatyh raikitvdgp reprrhrqkl ddqtkpgsls fserlseleq lrrtamrvsp 241 hhpaptpnpr aslnhstafn pqpqsqmqdt rqiqpsppws ydqsyqylgs iaspsvhpat 301 pispgrasgm ttlsaelssr lstapdltaf sdprqfpalp sisdprmhyp gaftysptpv 361 tsgigigmsa mgsatryhty lpppypgssq aqggpfqass psyhlyygas agsyqfsmvg 421 gerspprilp pctnastgsa llnpslpnqs dvveaegshs nsptnmapsa rleeavwrpy *** SEQ ID NO: 2 Human Runx1 (Isoform X1) AML1c LOCUS XP_054180869 480 aa linear PRI 20-MAR-2023 DEFINITION runt-related transcription factor 1 isoform X1 [Homo sapiens]. ACCESSION XP_054180869 VERSION XP_054180869.1 DBLINK BioProject: PRJNA807723 DBSOURCE REFSEQ: accession XM_054324894.1 KEYWORDS RefSeq. SOURCE Homo sapiens (human) Human Runx1 (Isoform X1) AML1c transcript sequence: 1 agatgggctg tggaaagggg aacagttaaa tttgtaattt gggttgtgtg aaaacttctt 61 tgggcctcat aaacaaccac agaaccacaa gttgggtagc ctggcagtgt cagaagtctg 121 aacccagcat agtggtcagc aggcaggacg aatcacactg aatgcaaacc acagggtttc 181 gcagcgtggt gagcatcacc aacccacagc caaggcggcg ctggcttttt tttttttttt 241 aatctttaac aatttgaata tttgttttta caaaggtgca ttttttaata gggcttgggg 301 agtcccagag gtatccagca ggggggagaa gaaagagaga tgtagggcta gaggggtgag 361 gctgaaacag tgacctgtct tggttttcgc tccgaaggta aaagaaatca ttgagtcccc 421 cgccttcaga agagggtgca ttttcaggag gaagcgatgg cttcagacag catatttgag 481 tcatttcctt cgtacccaca gtgcttcatg agagaatgca tacttggaat gaatccttct 541 agagacgtcc acgatgccag cacgagccgc cgcttcacgc cgccttccac cgcgctgagc 601 ccaggcaaga tgagcgaggc gttgccgctg ggcgccccgg acgccggcgc tgccctggcc 661 ggcaagctga ggagcggcga ccgcagcatg gtggaggtgc tggccgacca cccgggcgag 721 ctggtgcgca ccgacagccc caacttcctc tgctccgtgc tgcctacgca ctggcgctgc 781 aacaagaccc tgcccatcgc tttcaaggtg gtggccctag gggatgttcc agatggcact 841 ctggtcactg tgatggctgg caatgatgaa aactactcgg ctgagctgag aaatgctacc 901 gcagccatga agaaccaggt tgcaagattt aatgacctca ggtttgtcgg tcgaagtgga 961 agagggaaaa gcttcactct gaccatcact gtcttcacaa acccaccgca agtcgccacc 1021 taccacagag ccatcaaaat cacagtggat gggccccgag aacctcgaag acatcggcag 1081 aaactagatg atcagaccaa gcccgggagc ttgtcctttt ccgagcggct cagtgaactg 1141 gagcagctgc ggcgcacagc catgagggtc agcccacacc acccagcccc cacgcccaac 1201 cctcgtgcct ccctgaacca ctccactgcc tttaaccctc agcctcagag tcagatgcag 1261 gatacaaggc agatccaacc atccccaccg tggtcctacg atcagtccta ccaatacctg 1321 ggatccattg cctctccttc tgtgcaccca gcaacgccca tttcacctgg acgtgccagc 1381 ggcatgacaa ccctctctgc agaactttcc agtcgactct caacggcacc cgacctgaca 1441 gcgttcagcg acccgcgcca gttccccgcg ctgccctcca tctccgaccc ccgcatgcac 1501 tatccaggcg ccttcaccta ctccccgacg ccggtcacct cgggcatcgg catcggcatg 1561 tcggccatgg gctcggccac gcgctaccac acctacctgc cgccgcccta ccccggctcg 1621 tcgcaagcgc agggaggccc gttccaagcc agctcgccct cctaccacct gtactacggc 1681 gcctcggccg gctcctacca gttctccatg gtgggcggcg agcgctcgcc gccgcgcatc 1741 ctgccgccct gcaccaacgc ctccaccggc tccgcgctgc tcaaccccag cctcccgaac 1801 cagagcgacg tggtggaggc cgagggcagc cacagcaact cccccaccaa catggcgccc 1861 tccgcgcgcc tggaggaggc cgtgtggagg ccctactgag gcgccaggcc tggcccggct 1921 gggccacgcg ggccgccgcc ttcgcctccg ggcgcgcggg cctcctgttc gcgacaagcc 1981 cgccgggatc ccgggccctg ggcccggcca ccgtcctggg gccgagggcg cccgacggcc 2041 aggatctcgc tgtaggtcag gcccgcgcag cctcctgcgc ccagaagccc acgccgccgc 2101 cgtctgctgg cgccccggcc ctcgcggagg tgtccgaggc gacgcacctc gagggtgtcc 2161 gccggcccca gcacccaggg gacgcgctgg aaagcaaaca ggaagattcc cggagggaaa 2221 ctgtgaatgc ttctgattta gcaatgctgt gaataaaaag aaagatttta tacccttgac 2281 ttaacttttt aaccaagttg tttattccaa agagtgtgga attttggttg gggtgggggg 2341 agaggaggga tgcaactcgc cctgtttggc atctaattct tatttttaat ttttccgcac 2401 cttatcaatt gcaaaatgcg tatttgcatt tgggtggttt ttatttttat atacgtttat 2461 ataaatatat ataaattgag cttgcttctt tcttgctttg accatggaaa gaaatatgat 2521 tcccttttct ttaagtttta tttaactttt cttttggact tttgggtagt tgtttttttt 2581 tgttttgttt tgtttttttg agaaacagct acagctttgg gtcattttta actactgtat 2641 tcccacaagg aatccccaga tatttatgta tcttgatgtt cagacattta tgtgttgata 2701 attttttaat tatttaaatg tacttatatt aagaaaaata tcaagtacta cattttcttt 2761 tgttcttgat agtagccaaa gttaaatgta tcacattgaa gaaggctaga aaaaaagaat 2821 gagtaatgtg atcgcttggt tatccagaag tattgtttac attaaactcc ctttcatgtt 2881 aatcaaacaa gtgagtagct cacgcagcaa cgtttttaat aggattttta gacactgagg 2941 gtcactccaa ggatcagaag tatggaattt tctgccaggc tcaacaaggg tctcatatct 3001 aacttcctcc ttaaaacaga gaaggtcaat ctagttccag agggttgagg caggtgccaa 3061 taattacatc tttggagagg atttgatttc tgcccaggga tttgctcacc ccaaggtcat 3121 ctgataattt cacagatgct gtgtaacaga acacagccaa agtaaactgt gtaggggagc 3181 cacatttaca taggaaccaa atcaatgaat ttaggggtta cgattatagc aatttaaggg 3241 cccaccagaa gcaggcctcg aggagtcaat ttgcctctgt gtgcctcagt ggagacaagt 3301 gggaaaacat ggtcccacct gtgcgagacc ccctgtcctg tgctgctcac tcaacaacat 3361 ctttgtgttg ctttcaccag gctgagaccc taccctatgg ggtatatggg cttttacctg 3421 tgcaccagtg tgacaggaaa gattcatgtc actactgtcc gtggctacaa ttcaaaggta 3481 tccaatgtcg ctgtaaattt tatggcacta tttttattgg aggatttggt cagaatgcag 3541 ttgttgtaca actcataaat actaactgct gattttgaca catgtgtgct ccaaatgatc 3601 tggtggttat ttaacgtacc tcttaaaatt cgttgaaacg atttcaggtc aactctgaag 3661 agtatttgaa agcaggactt cagaacagtg tttgattttt attttataaa tttaagcatt 3721 caaattaggc aaatctttgg ctgcaggcag caaaaacagc tggacttatt taaaacaact 3781 tgtttttgag ttttcttata tatatattga ttatttgttt tacacacatg cagtagcact 3841 ttggtaagag ttaaagagta aagcagctta tgttgtcagg tcgttcttat ctagagaaga 3901 gctatagcag atctcggaca aactcagaat atattcactt tcatttttga caggattccc 3961 tccacaactc agtttcatat attattccgt attacatttt tgcagctaaa ttaccataaa 4021 atgtcagcaa atgtaaaaat ttaatttctg aaaagcacca ttagcccatt tcccccaaat 4081 taaacgtaaa tgtttttttt cagcacatgt taccatgtct gacctgcaaa aatgctggag 4141 aaaaatgaag gaaaaaatta tgtttttcag tttaattctg ttaactgaag atattccaac 4201 tcaaaaccag cctcatgctc tgattagata atcttttaca ttgaaccttt actctcaaag 4261 ccatgtgtgg agggggcttg tcactattgt aggctcactg gattggtcat ttagagtttc 4321 acagactctt accagcatat atagtattta attgtttcaa aaaaaatcaa actgtagttg 4381 ttttggcgat aggtctcacg caacacattt ttgtatgtgt gtgtgtgtgc gtgtgtgtgt 4441 gtgtgtgtga aaaattgcat tcattgactt caggtagatt aaggtatctt tttattcatt 4501 gccctcagga aagttaaggt atcaatgaga cccttaagcc aatcatgtaa taactgcatg 4561 tgtctggtcc aggagaagta ttgaataagc catttctact gcttactcat gtccctattt 4621 atgatttcaa catggataca tatttcagtt ctttcttttt ctcactatct gaaaatacat 4681 ttccctccct ctcttccccc caatatctcc ctttttttct ctcttcctct atcttccaaa 4741 ccccactttc tccctcctcc ttttcctgtg ttctcttaag cagatagcac atacccccac 4801 ccagtaccaa atttcagaac acaagaaggt ccagttcttc ccccttcaca taaaggaaca 4861 tggtttgtca gcctttctcc tgtttatggg tttcttccag cagaacagag acattgccaa 4921 ccatattgga tctgcttgct gtccaaacca gcaaactttc ctgggcaaat cacaatcagt 4981 gagtaaatag acagcctttc tgctgccttg ggtttctgtg cagataaaca gaaatgctct 5041 gattagaaag gaaatgaatg gttccactca aatgtcctgc aatttaggat tgcagatttc 5101 tgccttgaaa tacctgtttc tttgggacat tccgtcctga tgatttttat ttttgttggt 5161 ttttattttt ggggggaatg acatgtttgg gtcttttata catgaaaatt tgtttgacaa 5221 taatctcaca aaacatattt tacatctgaa caaaatgcct ttttgtttac cgtagcgtat 5281 acatttgttt tgggattttt gtgtgtttgt tgggaatttt gtttttagcc aggtcagtat 5341 tgatgaggct gatcatttgg ctcttttttt ccttccagaa gagttgcatc aacaaagtta 5401 attgtattta tgtatgtaaa tagattttaa gcttcattat aaaatattgt taatgcctat 5461 aacttttttt caattttttt gtgtgtgttt ctaaggactt tttcttaggt ttgctaaata 5521 ctgtagggaa aaaaatgctt ctttctactt tgtttatttt agactttaaa atgagctact 5581 tcttattcac ttttgtaaac agctaatagc atggttccaa ttttttttaa gttcactttt 5641 tttgttctag gggaaatgaa tgtgcaaaaa aagaaaaaga actgttggtt atttgtgtta 5701 ttctggatgt ataaaaatca atggaaaaaa ataaactttc aaattgaaat gacggtataa 5761 cacatctact gaaaaagcaa cgggaaatgt ggtcctattt aagccagccc ccacctaggg 5821 tctatttgtg tggcagttat tgggtttggt cacaaaacat cctgaaaatt cgtgcgtggg 5881 cttctttctc cctggtacaa acgtatggaa tgcttcttaa aggggaactg tcaagctggt 5941 gtcttcagcc agatgacatg agagaatatc ccagaaccct ctctccaagg tgtttctaga 6001 tagcacagga gagcaggcac tgcactgtcc acagtccatg gtacacagtc gggtgggccg 6061 cctcccctct cctgggagca ttcgtcgtgc ccagcctgag cagggcagct ggactgctgc 6121 tgttcaggag ccaccagagc cttcctctct ttgtaccaca gtttcttctg taaatccagt 6181 gttacaatca gtgtgaatgg caaataaaca gtttgacaag tacatacacc ata *** SEQ ID NO: 3 LOCUS NP_001104493 451 aa linear ROD 31-JUL-2023 DEFINITION runt-related transcription factor 1 isoform 3 [Mus musculus]. ACCESSION NP_001104493 VERSION NP_001104493.1 DBSOURCE REFSEQ: accession NM_001111023.2 KEYWORDS RefSeq. SOURCE Mus musculus (house mouse) Also known as AML1b Amino Acid Sequence of Mouse Runx1 (AML1b) ORIGIN 1 mripvdasts rrftppstal spgkmsealp lgapdggaal asklrsgdrs mvevladhpg 61 elvrtdspnf lcsvlpthwr cnktlpiafk vvalgdvpdg tlvtvmagnd enysaelrna 121 taamknqvar fndlrfvgrs grgksftlti tvftnppqva tyhraikitv dgpreprrhr 181 qklddqtkpg slsfserlse leqlrrtamr vsphhpaptp npraslnhst afnpqpqsqm 241 qdarqiqpsp pwsydqsyqy lgsitsssvh patpispgra sgmtslsael ssrlstapdl 301 tafgdprqfp tlpsisdprm hypgaftysp pvtsgigigm samssasryh tylpppypgs 361 sqaqagpfqt gspsyhlyyg asagsyqfsm vggersppri lppctnastg aallnpslps 421 qsdvvetegs hsnsptnmpp arleeavwrp y *** SEQ ID NO: 4 LOCUS NM_001111023 7138 bp mRNA linear ROD 31-JUL-2023 DEFINITION Mus musculus runt related transcription factor 1 (Runx1), transcript variant 3, mRNA. ACCESSION NM_001111023 VERSION NM_001111023.2 KEYWORDS RefSeq. SOURCE Mus musculus (house mouse) Also known as AML1b Transcript sequence of Mouse Runx1 (AML1b) ORIGIN 1 tctcattcat agagccagcg ggcgcgggca ggacgggcgc cccgcggccg gacccagcca 61 gggcaccacg ctgcccggcc ccgcggctgg cacttccatc ctgagctccc cgagacgcaa 121 gaaagtcacc ctttgcggct ctttctagat ctgcgttgga ccttcttgtt ttcctttcct 181 ttcctttttt ttttttttta atttttattt ttattttccc tttcttactt ttgaatgaac 241 cggcttctag gctttggatg tttcaacttc taggctgtga acttttctcc gattgtttct 301 tttgacaact gcgggagaaa gtgttccgga gttccaggcg agcggtgaag ttgcgtgtgc 361 gtggcagggt gcgtgggagt gagcgtgtgt aactcgagcc tactggtcta cggtgcctcc 421 tcccaggccc gctccacctg ctgcggttac acggtgctcg ctcgtgggta tttgtgcgcc 481 ccagcagaaa accagccacc gccgggcgac gttggaggaa gatcatcttg ccagactctg 541 ggctgactca ctcaaccaat gtagacagtg gctgacttcc gaagagtgcg agtctgcctg 601 tgtgtgtgac ccaactgcgg ctcaactccc accaaaccac aggatcagcc acaaacttaa 661 ccagcgttcc caaaccagag tcctcagctg tgggaagctc taccctgagt gtcatcgagt 721 ttgacttttt tttttttaag attagctaca gatctttgta aagcactgac catttatttg 781 gccagaattt attgagtaat gacaagtgat cctatcccag ccttccaact actgaagctg 841 attttcaagg ctacttaaaa aaaaaaaaac tgcagcaaac attaatggat ttctgttgtg 901 tttaaattct ctacagattg tattgtaaat attttatgaa gtagatcata tgtatatatt 961 tatatatacg agcacataca ttagtagcac aatcttttaa agttacggct cttgcttttg 1021 agaaacgaag tgagtttttc atggtaagag gggcgctctg tatggaagac actcctaagt 1081 tttgtatttt gttgagactc taaacaaaac tgacccgcaa gaaagaaaaa caaactgaca 1141 ggcaactaac tgctggaact tccaaatcta gtttttgctg gtcatttttt ggttgttgtt 1201 gttgttctgc gcataaatat tttaggacgc gtatgggaat tttgcctccg ggaccgtttg 1261 taatagccaa agactgaact tcaactctca aagcgaggct ctgttgggca tttgactttg 1321 aattctggtt gcccactttt taacgcacta agcggccagt tgctaaccct gcttgggtgt 1381 gaggccgatc cgtgtaacta gatctcagtc cgggactgat ggtaatttag tctcctaccc 1441 ccacaaccct ccggtagtaa taaaggcttc tgaacttgta tgttggtctc ccgggagcag 1501 cttgcagaag atccgagccc ctgtcgccgt ctagtaggag ctgttttcag ggtccttact 1561 caatcggctt gttgtgatgc gtatccccgt agatgccagc acgagccgcc gcttcacgcc 1621 gccttccacc gcgctgagcc ccggcaagat gagcgaggcg ctgccgctgg gcgccccgga 1681 tggcggcgcc gccctggcca gcaagctgag gagcggcgac cgcagcatgg tggaggtact 1741 agctgaccac cctggcgagc tagtgcgcac cgacagcccc aacttcctct gctccgtgct 1801 acccactcac tggcgctgca acaagaccct gcccatcgct ttcaaggtgg tggcactggg 1861 ggacgtcccg gatggcactc tggtcaccgt catggcaggc aacgatgaaa actactcggc 1921 agaactgaga aatgctaccg cggccatgaa gaaccaggta gcgagattca acgacctcag 1981 gtttgtcggg cggagcggta gaggcaagag cttcactctg accatcaccg tctttacaaa 2041 tccgccacaa gttgccacct accatagagc catcaaaatc acagtggacg gcccccgaga 2101 accccgaaga catcggcaga aactagatga tcagaccaag cccgggagtt tgtccttttc 2161 cgagcggctc agtgaattgg agcagctgcg gcgcacggcc atgagggtca gcccgcacca 2221 cccagccccc acgcccaacc ctcgggcctc cttgaaccac tccactgcct ttaaccctca 2281 gcctcaaagt cagatgcagg atgccaggca gatccagcca tccccaccgt ggtcctatga 2341 ccagtcctac cagtacctgg gatccatcac ctcttcctct gtccacccag cgacacccat 2401 ttcacccggc cgtgccagcg gcatgaccag cctctctgca gaactttcca gtcgactctc 2461 aacggctccg gacctgaccg ccttcggcga cccacgccag ttccctactc tgccgtccat 2521 ctccgacccg cgcatgcact acccaggcgc cttcacctac tcgccgcccg tcacgtcggg 2581 catcggcatc ggcatgtcag ccatgagctc ggcctctcgc taccacacct acctgccgcc 2641 gccctacccc ggctcatcac aggcgcaggc cgggcccttc cagaccggct cgccctccta 2701 ccatctatac tacggcgcct cggccggttc ctaccagttc tccatggtgg gcggagagag 2761 atcgcccccg cgcatcctgc cgccctgcac caacgcatcc accggcgccg cgctgctcaa 2821 ccccagcctc cccagccaga gcgacgtggt ggagaccgag ggcagccata gcaactcgcc 2881 caccaacatg ccccccgcgc gcctggagga ggccgtgtgg cggccctact gagctgagcg 2941 ccatcgccat cgagggactg ggcctgccgt ccatgcacag accccgccag gagggccctt 3001 ggaggccacc aggaagaatc ccggagggaa actgtgaatg cttctgattt agcaatgctg 3061 tgaataaaag aaagatttta tacccttgac ttcacttttt aaccacgttg tttattccaa 3121 agagtgtgga atgttttcgg ttcggggtgg ggaagacgca gcccatcctg tttggcatct 3181 atttcttatt tcggagtttt cttttccgca ccttatcgat tgcaaaaatg cctgtttgca 3241 tctgggtggt catttatttt taagtgtgta tagatttgag cttgcttttt tttcttcctt 3301 tgaccaactc aaagaaataa aattcccttc tctgtaaggt ttatttaact tttagacttt 3361 catgtagctg ggggttttat ttgtgtttgg tttttgtttt tatttttaaa gagacagcta 3421 cagctttggg tcatttttta actactgtat ttccacaaag aaatccctag atatttatgt 3481 atcttgatgt ttgaacattt acatatgtgt tgatactttt ttaattattt aaatgtactt 3541 atattaagaa agatatcaag tactacattt ttctttataa tagccaaagt taaatattat 3601 tgcgttgaag atgtctggaa aaaaaagaga tcgcttggtt aactagaaat attgtttaca 3661 ttaaactccc tttatgttat tcaaacaagt tggtaggtaa cgcagcaatg tttttaattg 3721 gattgtagac actgagggtc actccaaggt cagaagtaca aaattttctg ctaggctcaa 3781 caaatagtct catacctggc tccttccctt caaaaagaga ggcaaactct gtcctgaaag 3841 ggttcagaga ggtgccaagg atttgctctg aagaggattt cattttggcc tggagatata 3901 cttgccccaa ggcctcctca ttctggcatg ctttatcaca gagctcaacc aagtaagctg 3961 ttggtcaggg gtttacttac atagtattta catagaccca aaccactgaa tgtgattttt 4021 aaattgcctt ccattaatag tacccgttca ttgatgaaaa ccaaaacttg aggctgtacc 4081 ccaaagatcc aaatagaaga gttaagacca ggtgtctttg aggcctaaag gctgagtttt 4141 aagagagtgt accccaaaag tctgaaggag ccggtttcct tctcccagtc ttagtggaat 4201 cagtcatggg aggcagatgc cacgcccacc tgtgcaggat gctcctcaga agctgcccct 4261 tcaccagcat cttctcccac caggccgagc ccctgacctt tggggtgcat cagtgtgata 4321 gatcctggtc tctgcagtcc gccatggcta cggttcagat gtgcatcgtg tcactgtaaa 4381 tgtaatggta ctgttgttac agtggaggac ttggtcaaaa tccagttgtt ctacaacgta 4441 tgaagcctaa ccgctggttc tgacatacat gtgctcaaaa tgatctggtt gtttggattt 4501 ttcttttgtt gttttgtttt ttaatgtacc tcttaaatta gttgaagtga tgtcaggtca 4561 actccgaaga gcgtttgaaa gcaggacttc agcacagtgt ttgatttttt tattattatt 4621 aatattattt tataaattta agcattcaga ttagatcttt ggctgcaggc agcaaaaacg 4681 gctggactta tttaaaaaaa atacagcttg ttttttgagt tatctatatc tatatctata 4741 tgttgattct ttgtcttaca tagagcagca gcactttggt aacctgtgat accaggttgc 4801 tcttgtctgg agaagagcgc tagcaggatt cagagaaact cagaatagat cttcatatca 4861 gccatacctt cctcctccat ccggtctcca ctcagttatt ccacagaaca ctttgacagc 4921 tgtgttgtca gaaaaataaa aaaaaattta atttctcaaa aggagtttgt ttctccaaca 4981 ttagatgttc ctcttaccat aggctgccgt atctggcctg agaaaacggt agggaaggac 5041 gaaggaaaga gatttctatt ttttcatatt aattttgata tctaaagata cgctagccct 5101 cagaggagca gataatctca cacattgaat tttcgccctg ggcaccatgc atcaagaagg 5161 cttgtcactg tgttagagcc atttagtgct tcctaaactt ttatcaacat aggcagtatt 5221 tagtctcaga gaaaaaaaaa tccatcaggc acatgtagtc ttggagatag attccacggg 5281 gcaggtattt ctctacctga gaaattgtgt tcattgcctt cgggtgcttc cagcggtctc 5341 ctcattcgct gtcttcaagg aagacccata agccaattct gagataatgg agctgttggg 5401 aatactggtc cagagaaaga aaaatgggat aagccattct tactgcttat tcaagcccct 5461 atttataatt ttaacacact ttccattcct tctggttttc tcgccgtcta tatcctccca 5521 atagcccttc tcacttttct tttccctcct gcaaacacac acacacacac acacacacac 5581 acacataagg cacacacaca cacatcctct cccccatacc aagtgtccag aacacagaaa 5641 gtccagttct tctccgttta ttaaagaaca gggtgagtca gccattctct tgctcacggg 5701 tttttttccc caacagaaca gaggcgttgc cagccatttt gggtctgctt tctgtccaga 5761 tactgcagca aaaactcttg aggatcacaa cccgttggct gagcagctgt gctgctgccc 5821 aaacgtcctg cgcagacaaa cgcacgctgg gaccggaagg ggtgtctctc cttctgcctc 5881 ttttctttca tacgtttctc tcgaaaggcc tcaactgagg actgcaaatt tctttcttga 5941 aataactttc ccccagggac attcggtctt agggattttt tggttttgat gggttttgtt 6001 ttgttttggt ttttttggtt cttctcattt tctttgtagg agaaggcatg agatgttgag 6061 ggtctttcat acatgaaaat aaatagtttg acagcaatct cagaatatat tttttcctta 6121 tttgaacaaa gtactgtttt gtttactcta cagtacacct ttatttggtg ggtttggctg 6181 ttggtcggaa atgccttccc tctttcagcc aggtcggtat agccaaagct gatcatttgt 6241 ctgttttttc ttttttttcc tcagaagagt tgtatccaca aactcaattg tatttatgta 6301 tgtaaataga tttcatgctt cattataaaa tattgttaat gcctgtaata acttttttca 6361 cattttgtgt gtgtttctaa ggactttttc ttatgttcgc taaatattgt agaagaaaaa 6421 tgcttctctt aacttgttta ttttagattt taaaaaaacc aagctacttc ttactcactt 6481 ttataaacat aataaacaag catggttcca atgggttttt tttaagttca ccttatgttc 6541 taggagaaat gaatgtgcaa aaaaaaaaat caatcttcaa atgaactgtt gggtatttct 6601 gttactctgg atggaaaaag atcaatggaa aacaaaaaac aaaaaaacaa acttccaaat 6661 tgaaatagca gtaaaacaca tgtattgggg gtaaaaaaaa aaaaaaaaaa cacacctcag 6721 aaaatgcagt tctccagcct ccagctactg cccacacatg tggcaagtgt tagatgtggt 6781 cgtgaggaat cccaaaatca gtgcccgggc cttttccttc ctggtactaa cattaagaac 6841 acctcctaag gggcactgtc aggctcaggt cttaaagcac atagataaga gagaatgtcc 6901 cagaacgatt ctcagtagcc caggccagca ggtactcacg gcactgcacc tgctcactgc 6961 acacagtcgc tgaggcagac ccctctcctg ggggccatgc agagccaggg cagggcagcc 7021 ctaagctact gatgttttct ttcctttgta ctatggtctt ttctgtaact ccaatattca 7081 aacccatgtg aatggcaaat aaacaatttg acaagcacct aaaaaaaaaa aaaaaaaa *** SEQ ID NO: 5 LOCUS NP_001104491 465 aa linear ROD 31-JUL-2023 DEFINITION runt-related transcription factor 1 isoform 1 [Mus musculus]. ACCESSION NP_001104491 VERSION NP_001104491.1 DBSOURCE REFSEQ: accession NM_001111021.2 KEYWORDS RefSeq; RefSeq Select. SOURCE Mus musculus (house mouse) Longest isoform (AML1c) Transcript variant 1 This variant has different start site and different N- terminus compared to AML1b Amino acid sequence of Mouse Runx1 1 masdsifesf psypqcfmrd astsrrftpp stalspgkms ealplgapdg gaalasklrs 61 gdrsmvevla dhpgelvrtd spnflcsvlp thwrcnktlp iafkvvalgd vpdgtlvtvm 121 agndenysae lrnataamkn qvarfndlrf vgrsgrgksf tltitvftnp pqvatyhrai 181 kitvdgprep rrhrqklddq tkpgslsfse rlseleqlrr tamrvsphhp aptpnprasl 241 nhstafnpqp qsqmqdarqi qpsppwsydq syqylgsits ssvhpatpis pgrasgmtsl 301 saelssrlst apdltafgdp rqfptlpsis dprmhypgaf tysppvtsgi gigmsamssa 361 sryhtylppp ypgssqaqag pfqtgspsyh lyygasagsy qfsmvggers pprilppctn 421 astgaallnp slpsqsdvve tegshsnspt nmpparleea vwrpy *** SEQ ID NO: 6 LOCUS NM_001111021 5803 bp mRNA linear ROD 31-JUL-2023 DEFINITION Mus musculus runt related transcription factor 1 (Runx1), transcript variant 1, mRNA. ACCESSION NM_001111021 VERSION NM_001111021.2 KEYWORDS RefSeq; RefSeq Select. Transcript sequence ORIGIN 1 gtgaaaactt ctttggacct cataaacaac cacagaacca caagttgggt agcctggcag 61 tgtcagaagt gtaagcccag cacagtggtc agcaggcagg acgaatcaca ctgaatgcaa 121 accacaggct ttcgcagagc ggtgaaagaa attatagaat cccccgcctt caggagaggt 181 gcgttttcga aaggaaacga tggcttcaga cagcattttt gagtcatttc cttcatatcc 241 acagtgcttc atgagagatg ccagcacgag ccgccgcttc acgccgcctt ccaccgcgct 301 gagccccggc aagatgagcg aggcgctgcc gctgggcgcc ccggatggcg gcgccgccct 361 ggccagcaag ctgaggagcg gcgaccgcag catggtggag gtactagctg accaccctgg 421 cgagctagtg cgcaccgaca gccccaactt cctctgctcc gtgctaccca ctcactggcg 481 ctgcaacaag accctgccca tcgctttcaa ggtggtggca ctgggggacg tcccggatgg 541 cactctggtc accgtcatgg caggcaacga tgaaaactac tcggcagaac tgagaaatgc 601 taccgcggcc atgaagaacc aggtagcgag attcaacgac ctcaggtttg tcgggcggag 661 cggtagaggc aagagcttca ctctgaccat caccgtcttt acaaatccgc cacaagttgc 721 cacctaccat agagccatca aaatcacagt ggacggcccc cgagaacccc gaagacatcg 781 gcagaaacta gatgatcaga ccaagcccgg gagtttgtcc ttttccgagc ggctcagtga 841 attggagcag ctgcggcgca cggccatgag ggtcagcccg caccacccag cccccacgcc 901 caaccctcgg gcctccttga accactccac tgcctttaac cctcagcctc aaagtcagat 961 gcaggatgcc aggcagatcc agccatcccc accgtggtcc tatgaccagt cctaccagta 1021 cctgggatcc atcacctctt cctctgtcca cccagcgaca cccatttcac ccggccgtgc 1081 cagcggcatg accagcctct ctgcagaact ttccagtcga ctctcaacgg ctccggacct 1141 gaccgccttc ggcgacccac gccagttccc tactctgccg tccatctccg acccgcgcat 1201 gcactaccca ggcgccttca cctactcgcc gcccgtcacg tcgggcatcg gcatcggcat 1261 gtcagccatg agctcggcct ctcgctacca cacctacctg ccgccgccct accccggctc 1321 atcacaggcg caggccgggc ccttccagac cggctcgccc tcctaccatc tatactacgg 1381 cgcctcggcc ggttcctacc agttctccat ggtgggcgga gagagatcgc ccccgcgcat 1441 cctgccgccc tgcaccaacg catccaccgg cgccgcgctg ctcaacccca gcctccccag 1501 ccagagcgac gtggtggaga ccgagggcag ccatagcaac tcgcccacca acatgccccc 1561 cgcgcgcctg gaggaggccg tgtggcggcc ctactgagct gagcgccatc gccatcgagg 1621 gactgggcct gccgtccatg cacagacccc gccaggaggg cccttggagg ccaccaggaa 1681 gaatcccgga gggaaactgt gaatgcttct gatttagcaa tgctgtgaat aaaagaaaga 1741 ttttataccc ttgacttcac tttttaacca cgttgtttat tccaaagagt gtggaatgtt 1801 ttcggttcgg ggtggggaag acgcagccca tcctgtttgg catctatttc ttatttcgga 1861 gttttctttt ccgcacctta tcgattgcaa aaatgcctgt ttgcatctgg gtggtcattt 1921 atttttaagt gtgtatagat ttgagcttgc ttttttttct tcctttgacc aactcaaaga 1981 aataaaattc ccttctctgt aaggtttatt taacttttag actttcatgt agctgggggt 2041 tttatttgtg tttggttttt gtttttattt ttaaagagac agctacagct ttgggtcatt 2101 ttttaactac tgtatttcca caaagaaatc cctagatatt tatgtatctt gatgtttgaa 2161 catttacata tgtgttgata cttttttaat tatttaaatg tacttatatt aagaaagata 2221 tcaagtacta catttttctt tataatagcc aaagttaaat attattgcgt tgaagatgtc 2281 tggaaaaaaa agagatcgct tggttaacta gaaatattgt ttacattaaa ctccctttat 2341 gttattcaaa caagttggta ggtaacgcag caatgttttt aattggattg tagacactga 2401 gggtcactcc aaggtcagaa gtacaaaatt ttctgctagg ctcaacaaat agtctcatac 2461 ctggctcctt cccttcaaaa agagaggcaa actctgtcct gaaagggttc agagaggtgc 2521 caaggatttg ctctgaagag gatttcattt tggcctggag atatacttgc cccaaggcct 2581 cctcattctg gcatgcttta tcacagagct caaccaagta agctgttggt caggggttta 2641 cttacatagt atttacatag acccaaacca ctgaatgtga tttttaaatt gccttccatt 2701 aatagtaccc gttcattgat gaaaaccaaa acttgaggct gtaccccaaa gatccaaata 2761 gaagagttaa gaccaggtgt ctttgaggcc taaaggctga gttttaagag agtgtacccc 2821 aaaagtctga aggagccggt ttccttctcc cagtcttagt ggaatcagtc atgggaggca 2881 gatgccacgc ccacctgtgc aggatgctcc tcagaagctg ccccttcacc agcatcttct 2941 cccaccaggc cgagcccctg acctttgggg tgcatcagtg tgatagatcc tggtctctgc 3001 agtccgccat ggctacggtt cagatgtgca tcgtgtcact gtaaatgtaa tggtactgtt 3061 gttacagtgg aggacttggt caaaatccag ttgttctaca acgtatgaag cctaaccgct 3121 ggttctgaca tacatgtgct caaaatgatc tggttgtttg gatttttctt ttgttgtttt 3181 gttttttaat gtacctctta aattagttga agtgatgtca ggtcaactcc gaagagcgtt 3241 tgaaagcagg acttcagcac agtgtttgat ttttttatta ttattaatat tattttataa 3301 atttaagcat tcagattaga tctttggctg caggcagcaa aaacggctgg acttatttaa 3361 aaaaaataca gcttgttttt tgagttatct atatctatat ctatatgttg attctttgtc 3421 ttacatagag cagcagcact ttggtaacct gtgataccag gttgctcttg tctggagaag 3481 agcgctagca ggattcagag aaactcagaa tagatcttca tatcagccat accttcctcc 3541 tccatccggt ctccactcag ttattccaca gaacactttg acagctgtgt tgtcagaaaa 3601 ataaaaaaaa atttaatttc tcaaaaggag tttgtttctc caacattaga tgttcctctt 3661 accataggct gccgtatctg gcctgagaaa acggtaggga aggacgaagg aaagagattt 3721 ctattttttc atattaattt tgatatctaa agatacgcta gccctcagag gagcagataa 3781 tctcacacat tgaattttcg ccctgggcac catgcatcaa gaaggcttgt cactgtgtta 3841 gagccattta gtgcttccta aacttttatc aacataggca gtatttagtc tcagagaaaa 3901 aaaaatccat caggcacatg tagtcttgga gatagattcc acggggcagg tatttctcta 3961 cctgagaaat tgtgttcatt gccttcgggt gcttccagcg gtctcctcat tcgctgtctt 4021 caaggaagac ccataagcca attctgagat aatggagctg ttgggaatac tggtccagag 4081 aaagaaaaat gggataagcc attcttactg cttattcaag cccctattta taattttaac 4141 acactttcca ttccttctgg ttttctcgcc gtctatatcc tcccaatagc ccttctcact 4201 tttcttttcc ctcctgcaaa cacacacaca cacacacaca cacacacaca taaggcacac 4261 acacacacat cctctccccc ataccaagtg tccagaacac agaaagtcca gttcttctcc 4321 gtttattaaa gaacagggtg agtcagccat tctcttgctc acgggttttt ttccccaaca 4381 gaacagaggc gttgccagcc attttgggtc tgctttctgt ccagatactg cagcaaaaac 4441 tcttgaggat cacaacccgt tggctgagca gctgtgctgc tgcccaaacg tcctgcgcag 4501 acaaacgcac gctgggaccg gaaggggtgt ctctccttct gcctcttttc tttcatacgt 4561 ttctctcgaa aggcctcaac tgaggactgc aaatttcttt cttgaaataa ctttccccca 4621 gggacattcg gtcttaggga ttttttggtt ttgatgggtt ttgttttgtt ttggtttttt 4681 tggttcttct cattttcttt gtaggagaag gcatgagatg ttgagggtct ttcatacatg 4741 aaaataaata gtttgacagc aatctcagaa tatatttttt ccttatttga acaaagtact 4801 gttttgttta ctctacagta cacctttatt tggtgggttt ggctgttggt cggaaatgcc 4861 ttccctcttt cagccaggtc ggtatagcca aagctgatca tttgtctgtt ttttcttttt 4921 tttcctcaga agagttgtat ccacaaactc aattgtattt atgtatgtaa atagatttca 4981 tgcttcatta taaaatattg ttaatgcctg taataacttt tttcacattt tgtgtgtgtt 5041 tctaaggact ttttcttatg ttcgctaaat attgtagaag aaaaatgctt ctcttaactt 5101 gtttatttta gattttaaaa aaaccaagct acttcttact cacttttata aacataataa 5161 acaagcatgg ttccaatggg ttttttttaa gttcacctta tgttctagga gaaatgaatg 5221 tgcaaaaaaa aaaatcaatc ttcaaatgaa ctgttgggta tttctgttac tctggatgga 5281 aaaagatcaa tggaaaacaa aaaacaaaaa aacaaacttc caaattgaaa tagcagtaaa 5341 acacatgtat tgggggtaaa aaaaaaaaaa aaaaacacac ctcagaaaat gcagttctcc 5401 agcctccagc tactgcccac acatgtggca agtgttagat gtggtcgtga ggaatcccaa 5461 aatcagtgcc cgggcctttt ccttcctggt actaacatta agaacacctc ctaaggggca 5521 ctgtcaggct caggtcttaa agcacataga taagagagaa tgtcccagaa cgattctcag 5581 tagcccaggc cagcaggtac tcacggcact gcacctgctc actgcacaca gtcgctgagg 5641 cagacccctc tcctgggggc catgcagagc cagggcaggg cagccctaag ctactgatgt 5701 tttctttcct ttgtactatg gtcttttctg taactccaat attcaaaccc atgtgaatgg 5761 caaataaaca atttgacaag cacctaaaaa aaaaaaaaaa aaa *** SEQ ID NO: 7 Ad-mCherry-U6-n-Runx1-shRNA (virus number 2) Targeting Sequence: GCCCTCCTACCATCTATACTA *** SEQ ID NO: 8 Ad-mCherry-U6-n-Runx1-shRNA (virus number 2) shRNA sequence: 5’- CCGG-GCCCTCCTACCATCTATACTACTCGAGTAGTATAGATGGTAGGAGGGC-TTTTTG-3’ *** SEQ ID NO: 9 Ad-mCherry-U6-n-Runx1-shRNA (virus number 2) Scramble control shRNA sequence 5’ - CCGG-CAACAAGATGAAGAGCACCAACTCGAGTTGGTGCTCTTCATCTTGTTG-TTTTT -3’ *** SEQ ID NO: 10 Runx1 shRNA short hairpin forward primer sequence: 5^F@SBBBBFFFBBBSBBS@BB@SBS@S@BS@BSBF@FS@FS@S@F@SFFS@FF@FFFB TTTTTGG-3^ *** SEQ ID NO: 11 Runx1 shRNA short hairpin reverse primer sequence: 5^@@SSBB@@@@@FBBBSBBS@BB@SBS@S@BS@BSBF@FS@FS@S@F@SFFS@FF@FFF CCCGGG-3^ *** SEQ ID NO: 12 Scramble shRNA short hairpin forward primer sequence: 5^F@SBBBBFFB@@B@@F@SF@@F@FB@BB@@BSBF@FSSFFSFBSBSSB@SBSSFSSFS TTTTG-3^ *** SEQ ID NO: 13 Scramble shRNA short hairpin reverse primer sequence: 5^@@SSB@@@@@B@@B@@F@SF@@F@FB@BB@@BSBF@FSSFFSFBSBSSB@SBSSFSSF CCGGG-3^ *** SEQ ID NO: 14 pZacf-U6-luc-ZsGreen Full sequence: ctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtcgggcgacctttggtcgcccggcctcagtgagcgagcgagcg cgcagagagggagtggccaactccatcactaggggttccttgtagttaatgattaacccgccatgctacttatctacgtagccatgctctaggaa gatctgggcaggaagagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagagataattagaattaatttgactgta aacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatg cttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaaggacgaggatcccgtgcgttgctagtaccaacttcaagagagttg gtactagcaacgcacttttttacgcgtgaattctagttattaatagtaatcaattacggggtcattagttcatagcccatatatggagttccgcgttaca taacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaata gggactttccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctatt gacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcg ctattaccatggtgatgcggttttggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtca atgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtaggcgtgtacg gtgggaggtctatataagcagagctggtttagtgaaccgtcagatccgctagcgctaccggtcgccaccatggcccagtccaagcacggcct gaccaaggagatgaccatgaagtaccgcatggagggctgcgtggacggccacaagttcgtgatcaccggcgagggcatcggctacccctt caagggcaagcaggccatcaacctgtgcgtggtggagggcggccccttgcccttcgccgaggacatcttgtccgccgccttcatgtacggca accgcgtgttcaccgagtacccccaggacatcgtcgactacttcaagaactcctgccccgccggctacacctgggaccgctccttcctgttcga ggacggcgccgtgtgcatctgcaacgccgacatcaccgtgagcgtggaggagaactgcatgtaccacgagtccaagttctacggcgtgaac ttccccgccgacggccccgtgatgaagaagatgaccgacaactgggagccctcctgcgagaagatcatccccgtgcccaagcagggcatc ttgaagggcgacgtgagcatgtacctgctgctgaaggacggtggccgcttgcgctgccagttcgacaccgtgtacaaggccaagtccgtgcc ccgcaagatgcccgactggcacttcatccagcacaagctgacccgcgaggaccgcagcgacgccaagaaccagaagtggcacctgacc gagcacgccatcgcctccggctccgccttgccctaactcgactagagtcgacccgggcggccgcttcgagcagacatgataagatacattga tgagtttggacaaaccacaactagaatgcagtgaaaaaaatgctttatttgtgaaatttgtgatgctattgctttatttgtaaccattataagctgcaa taaacaagttaacaacaacaattgcattcattttatgtttcaggttcagggggagatgtgggaggttttttaaagcaagtaaaacctctacaaatgt ggtaaaatcgataaggatcttcctagagcatggctacgtagataagtagcatggcgggttaatcattaactacaaggaacccctagtgatgga gttggccactccctctctgcgcgctcgctcgctcactgaggccgggcgaccaaaggtcgcccgacgcccgggctttgcccgggcggcctcag tgagcgagcgagcgcgcagccttaattaacctaattcactggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaat cgccttgcagcacatccccctttcgccagctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaatgg cgaatgggacgcgccctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcg cccgctcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagggttccgatttagtgct ttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagt ccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggcct attggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaaatattaacgcttacaatttaggtggcacttttcggggaaatgtg cgcggaacccctatttgtttatttttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttcaataatattgaaaaagg aagagtatgagtattcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaa agatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgt tttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactatt ctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccat gagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaac tcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttg cgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgc gctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggt aagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcact gattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttt tgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatccttttt ttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaagg taactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctaca tacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataag gcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtga gctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgag ggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcagggggg cggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgttctttcctgcgttatcccctgatt ctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagccgaacgaccgagcgcagcgagtcagtgagcgaggaagc ggaagagcgcccaatacgcaaaccgcctctccccgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcg ggcagtgagcgcaacgcaattaatgtgagttagctcactcattaggcaccccaggctttacactttatgcttccggctcgtatgttgtgtggaattgt gagcggataacaatttcacacaggaaacagctatgaccatgattacgccagatttaattaaggccttaattagg Detailed Description of the Invention The present invention will now be discussed with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this text are incorporated herein by reference. The features disclosed in the foregoing description, or in the following claims, or in the accompanying drawings, expressed in their specific forms or in terms of a means for performing the disclosed function, or a method or process for obtaining the disclosed results, as appropriate, may, separately, or in any combination of such features, be utilised for realising the invention in diverse forms thereof. While the invention has been described in conjunction with the exemplary embodiments described herein, many equivalent modifications and variations will be apparent to those skilled in the art when given this disclosure. Accordingly, the exemplary embodiments of the invention set forth herein are considered to be illustrative and not limiting. Various changes to the described embodiments may be made without departing from the spirit and scope of the invention. For the avoidance of any doubt, any theoretical explanations provided herein are provided for the purposes of improving the understanding of a reader. The inventors do not wish to be bound by any of these theoretical explanations. Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Throughout this specification, including the claims which follow, unless the context requires kpdanseoa, pda skn` ^_kilneoa^ ]j` ^ej_hq`a^, ]j` r]ne]pekjo oq_d ]o ^_kilneoao^, ^_kilneoejc^, ]j` ^ej_hq`ejc^ sehh ^a qj`anopkk` pk eilhu pda ej_hqoekj kb ] op]ted integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps. It must be noted that, as used in the specification and the appended claims, the singular bknio ^],^ ^]j,^ ]j` ^pda^ ej_hq`a lhqn]h nabanajpo qjhaoo pda _kjpatp _ha]nhu `e_p]pao kpdanseoa. Q]jcao i]u ^a atlnaooa` danaej ]o bnki ^]^kqp^ kja l]npe_qh]n r]hqa, and / or to ^]^kqp^ ]jkpdan l]npe_qh]n r]hqa. Vdaj oq_d ] n]jca eo atlnaooa`, ]jkpdan ai^k`eiajp includes from the one particular value and / or to the other particular value. Similarly, when r]hqao ]na atlnaooa` ]o ]llnktei]pekjo, ^u pda qoa kb pda ]jpa_a`ajp ^]^kqp,^ ep sehh ^a qj`anopkk` pd]p pda l]npe_qh]n r]hqa bknio ]jkpdan ai^k`eiajp. Sda pani ^]^kqp^ ej nah]pekj to a numerical value is optional and means for example + / - 10%. Examples Examples are provided below to facilitate a more complete understanding of the invention. The following examples illustrate the exemplary modes of making and practicing the invention. However, the scope of the invention is not limited to specific embodiments disclosed in these Examples, which are for the purposes of illustration only. Example 1 - Materials and Methods Animals The care and use of animals were in accordance with the UK Government Animals (Scientific Procedures) Act 1986 and all procedures were approved by the University of Glasgow Animal Welfare and Ethical Review Body and licensed by the Home Office, UK (project licence no. P05FEIF82, expired April 2023 and PP4465428, expires March 2028). Male mice (10-14 weeks, 25-30g) were singly housed on a 12 / 12 h light / dark cycle and fed and watered ad libitum and were randomly assigned to experimental groups. Animals were euthanised by schedule one procedures (cervical dislocation). Two-hit HFpEF Model Hypertension was induced by giving mice N^-Nitro-L-arginine methyl ester hydrochloride (L- NAME, Sigma-N5751) in their drinking water resulting in increased blood pressure. Control mice were supplied with tap water. High-fat diet (Research diet-D12492) with fat (60% Kcal / 100 grams) was used to induce weight gain and metabolic distress, while control mice were fed a standard chow diet (SDS-801151) with fat (7.37% Kcal / 100 grams). Body weight and blood pressure were monitored weekly. Generation of cardiomyocyte-specific Runx1-deficient mice Runx1-deficient (Runx1^ / ^) mice were generated as previously described (Tsao, C.W. et al. Temporal trends in the incidence of and mortality associated with heart failure with preserved and reduced ejection fraction. JACC Hear. Fail.6, 678~685 (2018)). Briefly, mice with floxed Runx1 (Runx1fl / fl, The Jackson Laboratory, ME, USA) were crossed with mice expressing tamoxifen-inducible Cre recombinase (MerCreMer) under the control of the cardiac-ola_ebe_ ^LGB (^-myosin heavy chain) to generate the relevant test (Runx1^ / ^) and littermate (Runx1fl / fl) control groups. Echocardiography Mice were anaesthetised in a pre-filled induction chamber with 5% isoflurane in 1.0 L / min O2and transferred to a face mask and maintained at 0.5-1%. Echocardiography was performed using a Siemens ACUSON Seqoula C512 and a 15L8 Transducer and a VEVOF2 Imaging System from Visual Sonics and a 30MHz transducer. Pressure Volume Loop (PVL) measurements Pressure~volume (PV) loop measurements were recorded using the Transonic ADV500 small animal model PV measurement system and a 1.2 F 3.5 mm spaced PV admittance catheter (FTH-1212B-4517, Scisense, UK). Mice were anesthetized in a pre-filled induction chamber with 5% isofluorane gas (Isoflo, Abbott Laboratories, USA) delivered in O2 at 1.5 L / min. Isofluorane concentration was gradually reduced to 1.5-2% during the procedure and maintained at this level for the duration of the measurements. A longitudinal incision was made in the centre of the neck and the right carotid artery dissected free of the surrounding tissue. A small incision was made in the vessel and the catheter inserted into the vessel and advanced into the left ventricle of the heart. Steady state readings were then taken for 15 min. Data were recorded in the Labchart software via an eight-channel PowerLab. Blood Pressure Measurements BP-2000 Blood Pressure Analysis Systemz (BIOSEBLAB instruments-BP-2000) was used for tail-cuff mice blood pressure measurements. Before beginning the study, animals were familiarized with the heating chamber, the restraint and tail cuff application process before the actual blood pressure measurement session. Also, 2-3 training sessions were performed prior to commencing actual readings. The animals were handled calmly and confidently during the procedure since stress and excess movement will affect the blood pressure recording. Animals were acclimatized for 10 mins to the equipment before using any readings and minimum of 3 separate measuring sessions were conducted over a 3 to 5- day period. Following acclimatization, measurements were taken for each mouse. Exercise Intolerance Testing For the exercise intolerance testing, a Forced Exercise Walking Wheel (model 80800A; PsymCon control device model (35500); Lafayette instruments) was used. Mice were acclimatised to the exercise wheel for three days before exercise intolerance test. For acclimatisation, mice were placed inside the stationary exercise wheel for 10 min on the first day, then were placed for 5 min at 3 m / min on the second day, and 4 m / min for 10 min on the third day. For intolerance test, animals began with 3 m / min warm-up speed for 5 min followed by an increase to 4 m / min until they became intolerant. Exercise intolerance is defined as the inability of the animal to return to running within 10 sec of coasting inside the wheel. Histology Mice were anaesthetised in an induction chamber at 4% isoflurane and once righting reflex was lost were sacrificed by cervical dislocation. Hearts were quickly removed, flushed with saline, and fixed in 10% buffered formalin for 24h before being embedded in a wax block for sectioning. Fibrosis was characterised via picrosirius Red-stained histological sections imaged at 20x magnification on a Leica Aperio AT2 slide scanner. Images were analysed for each heart separately by taking 4 random sections from different sites of left ventricular tissue (apex, posterior wall, anterior wall), then analysed using ImageJ software. AAV-mediated knockdown of Runx1 shRNA sequences for murine Runx1 were manufactured by the short hairpin sequences in the adenoviral vectors for Runx1 and scramble. Mice were intravenously injected via the tail vein with either AAV9-scramble-shRNA or AAV9-Runx1-shRNA (1x1010virus particles per mouse) in a randomised fashion. shRNA sequences targeting murine Runx1 were generated using the short hairpin sequences in the adenoviral vectors for both Runx1 and scramble. Briefly, the complementary oligos of shRNA (Eurofins genomics, Germany) were annealed and cloned into a pZac2.1 vector, which was then used to produced recombinant AAV vectors in the AAV Vector Unit at IGGEB Triest. Viral stocks were obtained by PEG precipitation and two subsequent CsCl2 Gradient centrifugations. Titration of AAV viral particles was performed by real-time PCR quantification of the number of packaged viral genomes. Runx1 shRNA encoded Runx1 shRNA short hairpin forward: 5^F@SBBBBFFFBBBSBBS@BB@SBS@S@BS@BSBF@FS@FS@S@F@SFFS@FF@FFFB TTTTTGG-3^ [SEQ ID NO: 10] and reverse 5^@@SSBB@@@@@FBBBSBBS@BB@SBS@S@BS@BSBF@FS@FS@S@F@SFFS@FF@FFF CCCGGG-3^ [SEQ ID NO: 11]. Scramble shRNA short hairpin contained forward: 5^F@SBBBBFFB@@B@@F@SF@@F@FB@BB@@BSBF@FSSFFSFBSBSSB@SBSSFSSFS TTTTG-3^ [SEQ ID NO: 12] and reverse 5^@@SSB@@@@@B@@B@@F@SF@@F@FB@BB@@BSBF@FSSFFSFBSBSSB@SBSSFSSF CCGGG-3^ [SEQ ID NO: 13]. Ro5-3335 knockdown of Runx1 C57BL / 6J mice were randomly assigned to receive either vehicle (100% DMSO) or 20 mg / kg of Ro5-3335 (Tocris-Bioscience, UK) given subcutaneously every second day in the final two weeks of the HFpEF protocol for a total of 7 injections. Ro24-7429 knockdown of Runx1 Experiments for Ro24-7429 were conducted as for Ro5-3335. Statistics Data were expressed as mean ± SEM. Comparisons between two experimental groups sana lanbknia` sepd pda Rpq`ajp^o p-test on raw data before normalization to percentage change where appropriate. Comparisons between more than two groups were conducted on raw data with ANOVA. Example 2 - Effect of Runx1-deficiency on the development of HFpEF We used a previously established (Schiattarella, G.G. et al. Nitrosative stress drives heart failure with preserved ejection fraction. Nature 568, 351~356 (2019)), two-hit model (2HM) that combines administration of a high-fat diet (HFD) and inhibition of nitric oxide synthase with N^-nitro-L-arginine methyl ester (L-NAME) in drinking water to induce a HFpEF phenotype and compared changes to age-matched controls (CTRL). To evaluate the contribution of RUNX1 to the pathophysiology of HFpEF, we utilised cardiomyocyte-specific Runx1-deficient (Runx1^ / ^) mice and floxed control mice (Runx1fl / fl) on this protocol (Figure 1a). After 10-12 weeks, each of the two-hits were observed in 2HM- Runx1fl / fland 2HM- Runx1^ / ^mice with an increase in body weight (10% and 8%, respectively p < 0.05; Figure 1b) and systolic blood pressure (SBP, 17% and 14% respectively, p < 0.05; Figure 1c) compared to CTRL-Runx1fl / fland CTRL-Runx1^ / ^mice. We then evaluated multiple hallmark parameters of the HFpEF phenotype. At the end of the 12-week protocol 2HM-Runx1fl / flmice had developed exercise intolerance, demonstrated by 56% reduction in running distance compared to CTRL-Runx1fl / fl(p = 0.003; Figure 1d). 2HM-Runx1^ / ^mice also had reduction in running distance compared to control (41%, p = 0.003; Figure 1d) however the exercise intolerance was attenuated because they ran greater distances than 2HM-Runx1fl / flmice (128 ± 13m vs.192 ± 13m, p = 0.0216; Figure 1d). Runx1 deficiency also protected mice from development of pulmonary oedema. Wet to dry lung weight ratio was increased in 2HM-Runx1fl / flcompared to in CTRL-Runx1fl / flmice (3.90 ± 0.2 vs.2.84 ± 0.2, p = 0.0024; Figure 1e). Conversely, there was no difference between 2HM-Runx1^ / ^and CTRL-Runx1^ / ^mice (3.00 ± 0.1, p = 0.9686; Figure 1e), had significantly lower wet to dry lung weight ratio than Runx1fl / fl(2HM) mice (3.14 ± 0.2 vs.2.84 ± 0.2, p = 0.0346; Figure 1e). Runx1 deficiency was also protective against development of hypertrophy as measured by LV weight normalised to tibial length (LV / TL) and cardiomyocyte cross sectional area (CSA). 2HM-Runx1fl / flhad increased LV / TL compared to CTRL-Runx1fl / flmice (4.9 ± 2.3 *10-3vs.3.5 ± 1.2 *10-3, p < 0.0001; Figure 1f) and 2HM- Runx1^ / ^mice had smaller LV / TL than 2HM-Runx1fl / flmice (4.2 ± 1.3 *10-3, p = 0.0198; Figure 1f) but no difference compared to CTRL-Runx1^ / ^mice (3.7 ± 1.0 *10-3, p = 0.1225; Figure 1f). This pattern was also demonstrated with cardiomyocyte cross sectional area (CTRL-Runx1fl / fl: 352 { 9.9 ^i2, CTRL-Runx1^ / ^: 323 { 32.4 ^i2, 2HM-Runx1fl / fl: 482 ± 18.0 ^i2, 2HM-Runx1^ / ^: 289 { 24.8 ^i2; Figure 1g). Additionally, there is a striking preservation of diastolic function as a result of cardiomyocyte-specific Runx1 knockdown, quantified by E to A wave ratio from M-mode echocardiography and the end-diastolic pressure-volume relationship (EDPVR), measured during vena cava occlusion during pressure-volume loop surgery. Compared to CTRL-Runx1fl / fland CTRL-Runx1^ / ^, 2HM-Runx1fl / flhad a higher E / A ratio whereas 2HM-Runx1^ / ^mice did not, and in fact also had a lower E / A ratio than 2HM- Runx1fl / flmice (CTRL-Runx1fl / fl: 1.63 ± 0.10, CTRL-Runx1^ / ^: 1.58 ± 0.09, 2HM-Runx1fl / fl: 3.75 ± 0.29, 2HM-Runx1^ / ^: 1.67 ± 0.09; Figure 1h). An independent measure of LV chamber stiffness was calculated by fitting the slope of the EDPVR. 2HM-Runx1fl / flmice had a steeper EDPVR slope than 2HM-Runx1^ / ^mice, indicating better diastolic function (0.075 ± 0.008 vs.0.018 ± 0.003, p < 0.0001; Figure 1i). Example 3 - Runx1 RNA interference attenuates diastolic dysfunction Given the striking phenotypic differences observed in 2HM-Runx1^ / ^mice we next tested whether using translational approaches to target RUNX1 expression could prevent the development of HFpEF. We utilised a viral vector mediated gene delivery approach with AAV9-Runx1-shRNA to knockdown Runx1 in our 2HM of HFpEF. We injected 12-week-old C57BL / 6N mice with AAV9-scramble-shRNA (2HM-AAV9-scram, n = 10) or AAV9-Runx1-shRNA (2HM-AAV9- Runx1, n = 11) after which we delivered the 2HM protocol for 8 weeks and compared them to aged-matched controls C57BL / 6N mice and on the 2HM (2HM-C57N) and control (CTRL- C57N, Figure 2a). The study began by once again confirming the efficacy of our 2HM by measuring changes in body weight (Figure 2b) and SBP (Figure 2c) over the duration of the protocol in all three 2HM groups compared to the CTRL group (Change in body weight ~ CTRL-C57N: 2.8 ± 0.7g, 2HM-C57N: 8.0 ± 0.8g, 2HM-AAV9-scram: 9.2 ± 1.2g, 2HM-AAV9-Runx1: 10.1 ± 1.0g; change in SBP ~ CTRL-C57N: -15.2 ± 12.5mmHg, 2HM-C57N: 20.3 ± 3.1mmHg, 2HM- AAV9-scram: 23.6 ± 6.2mmHg, 2HM-AAV9-Runx1: 27.2 ± 3.7mmHg). Targeting Runx1 with AAV9 was effective in preventing some, but not all, of the HFpEF symptoms we characterised. Exercise intolerance was observed in all three 2HM groups compared to CTRL-C57N, but with no difference in running distance between them (CTRL- C57N: 262 ± 12m, 2HM-C57N: 108 ± 3m, 2HM-AAV9-scram: 108 ± 23m, 2HM-AAV9- Runx1: 163 ± 22m; Figure 2d). This was also consistent for LV / TL (CTRL-C57N: 4.1 ± 0.2 *10-3, 2HM-C57N: 5.0 ± 0.2 *10-3, 2HM-AAV9-scram: 5.4 ± 0.2 *10-3, 2HM-AAV9-Runx1: 4.9 ± 0.2 *10-3; Figure 2f). However, although cardiomyocyte cross sectional area was increased in both 2HM-AAV9 groups compared to CTRL-C57N, it was reduced in 2HM- AAV9-scram compared to 2HM-AAV9-Runx1 (CTRL-B57M: 217 { 20 ^i2, 2HM-AAV9- o_n]i: 449 { 10 ^i2, 2HM-AAV9-Runx1: 308 { 23 ^i2; Figure 2g). AAV9-Runx1 also attenuated the development pulmonary oedema compared to the other two 2HM groups, quantified by wet to dry lung weight ratio (CTRL-C57N: 3.18 ± 0.16, 2HM-C57N: 4.58 ± 0.07, 2HM-AAV9-scram: 4.36 ± 0.12, 2HM-AAV9-Runx1: 3.68 ± 0.15; Figure 2e). Most striking was the preservation of diastolic function by targeting Runx1 with AAV9. E / A ratio was increased in both 2HM-C57N and 2HM-AAV9-scram groups compared to CTRL-C57N, but was not increased in 2HM-AAV9-Runx1 compared to CTRL-C57N (CTRL-C57N: 1.32 ± 0.10, 2HM-C57N: 2.55 ± 0.26, 2HM-AAV9-scram: 2.78 ± 0.33, 2HM-AAV9-Runx1: 1.29 ± 0.12; Figure 2h, 2i). This was also consistent with EDPVR, which was lower in the 2HM- AAV9-Runx1 group compared to 2HM-AAV9-scram (0.077 ± 0.009 vs.0.019 ± 0.003, p < 0.0001; Figure 2j). Example 4 - Small molecule inhibition of Runx1 remedies the HFpEF phenotype To take this one step further, we aimed to identify if inhibition of RUNX1 could ameliorate the HFpEF phenotype once it has already begun to develop. 12-week-old C57BL / 6N strain mice were placed on the 2HM protocol for 10-12 weeks. Prior to treatment, in vivo parameters were utilised to ensure the HFpEF phenotype had developed and any mice that did not have HFpEF symptoms were excluded. We then injected a small molecule inhibitor of RUNX1, Ro5-3335, every second day for two weeks prior to collecting end-point measurements and organometrics (Figure 3a). Although Ro5-3335 injections did not change exercise tolerance (Figure 3b) or hypertrophy (Figure 3d), pulmonary oedema was less severe in 2HM-Ro5-3335 mice compared to 2HM- DMSO mice (4.26 ± 0.05 vs.4.06 ± 0.07, p = 0.0313; Figure 3c). Although two weeks of Ro5-3335 injections does not remarkably change the HFpEF phenotype overall, the preservation of diastolic function is striking in 2HM-Ro5-3335 mice compared to 2HM- DMSO. There is no difference in E / A wave ratio post-injection compared to pre-injection in the 2HM-DMSO mice (2.85 ± 0.44 vs.2.93 ± 0.18, p = 0.8810; Figure 3e, left) whereas the post-injection E / A wave ratio was reduced in the 2HM-Ro5-3335 mice compared to pre- injection, demonstrating a recovery of diastolic function (1.68 ± 0.14 vs.2.51 ± 0.16, p = 0.0063; Figure 3e, right). PV loop assessment of diastolic function by EDPVR also demonstrated diastolic dysfunction in 2HM-DMSO mice compared to the 2HM-Ro5-3335 group (0.045 ± 0.005 vs.0.029 ± 0.005, p = 0.048; Figure 3f). Similar results were found for another small molecule inhibitor of RUNX1, Ro24-7429. Experiments were carried out exactly as for Ro5-3335. The results are presented in Figure 4. Example 5 - Discussion The experimental results described herein demonstrate an important role for RUNX1 in the development of HFpEF. The contribution of Runx1 in the development of a HFpEF phenotype is demonstrated using a cardiomyocyte-specific Runx1 deficient mouse model. Further, the experiments demonstrate that targeting Runx1 in the context of HFpEF shows therapeutic potential for the prevention and / or treatment of HFpEF. In particular, small molecule inhibitors have been found with therapeutic potential for the prevention and / or treatment of HFpEF. References A number of publications are cited above in order to more fully describe and disclose the invention and the state of the art to which the invention pertains. Full citations for these references are provided below. The entirety of each of these references is incorporated herein. Tsao, C. W. et al. Temporal trends in the incidence of and mortality associated with heart failure with preserved and reduced ejection fraction. JACC Hear. Fail.6, 678~685 (2018). Omote, K., Verbrugge, F.H. & Borlaug, B.A. Heart failure with preserved ejection fraction: mechanisms and treatment strategies. Annu. Rev. Med.73, 321~337 (2022). Mishra, S. & Kass, D.A. Cellular and molecular pathobiology of heart failure with preserved ejection fraction. Nat. Rev. Cardiol.18, 400~423 (2021). McCarroll, C.S. et al. Runx1 deficiency protects against adverse cardiac remodeling after myocardial infarction. Circulation 137, 57~70 (2018). Riddell, A. et al. RUNX1: An emerging therapeutic target for cardiovascular disease. Cardiovasc. Res.116, 1410~1423 (2020). Schiattarella, G.G. et al. Nitrosative stress drives heart failure with preserved ejection fraction. Nature 568, 351~356 (2019). For standard molecular biology techniques, see Sambrook, J., Russel, D.W. Molecular Cloning, A Laboratory Manual.3 ed.2001, Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.

Claims

Claims 1. A method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), in a patient having or at risk of developing HFpEF, wherein the method comprises administering to the patient a therapeutically effective amount of an agent that decreases the expression or activity of Runx1 in the patient.

2. A method according to claim 1, wherein Runx1 has an amino acid sequence of SEQ ID NO:

1.

3. A method according to claim 1 or claim 2, wherein the activity or expression of Runx1 is decreased by at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or by 100% compared to a baseline value or a control value.

4. A method according to any preceding claim, wherein the decrease in activity or expression of Runx1 is confined to cardiomyocytes.

5. A method according to any preceding claim, wherein the agent is a Runx1 inhibitor.

6. A method according to claim 5, wherein the Runx1 inhibitor is a:, wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl,R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof.

7. A method according to claim 5 or claim 6, wherein the Runx1 inhibitor is:wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3).

8. A method according to any of claims 5 to 7, wherein the Runx1 inhibitor is:wherein X is N(R,R1) or O R and R1are each independently C1-C4 alkyl or wherein one of R and R1 is hydrogen, preferably wherein N(R,R1) is NH(CH3).

9. A method according to any of claims 5 to 8, wherein the inhibitor is:

10. A method according to any of claims 5 to 9, wherein the Runx1 inhibitor is Ro5-3335 or Ro24- 7429, or a combination thereof.

11. A composition comprising an agent that decreases the expression or activity of Runx1 for use in a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), in a patient having or at risk of developing HFpEF, wherein the method comprises administering a therapeutically effective amount of the composition to the subject.

12. A composition for the use according to claim 11, wherein Runx1 has an amino acid sequence of SEQ ID NO:

1.

13. A composition for the use according to claim 11 or claim 12, wherein the activity or expression of Runx1 is decreased by at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or by 100% compared to a baseline value or a control value.

14. A composition for the use according to any of claims 11 to 13, wherein the agent that decreases the expression or activity of Runx1 is a Runx1 inhibitor.

15. A composition for the use according to claim 14, wherein the Runx1 inhibitor is:, wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof.

16. A composition for the use according to claim 14 or claim 15, wherein the Runx1 inhibitor is:wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3), 17. A composition for the use according to any of claims 14 to 16, wherein the Runx1 inhibitor is:wherein X is N(R,R1) or O R and R1are each independently C1-C4 alkyl or wherein one of R and R1 is hydrogen, preferably wherein N(R,R1) is NH(CH3)18. A composition for the use according to any of claims 14 to 17, wherein the Runx1 inhibitor is:

19. A composition for the use according to any of claims 14 to 18, wherein the Runx1 inhibitor is Ro5- 3335 or Ro24-7429, or a combination thereof.

20. A composition for the use according to any of claims 11 to 13, wherein the decrease in activity or expression of Runx1 is confined to cardiomyocytes.

21. A composition for the use according to any of claims 11 to 13 or claim 20, wherein the agent that decreases the expression or activity of Runx1 is a nucleic acid that is configured to target Runx1.

22. A composition for the use according to claim 21, wherein the agent that decreases the expression or activity of Runx1 is a nucleic acid that specifically binds to a nucleic acid encoding Runx1.

23. A composition for the use according to claim 21 or 22, wherein the nucleic acid encoding Runx1 has a sequence of SEQ ID NO:

2.

24. A composition for the use according to any of claims 21 to 23, wherein the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is expressed only in cardiomyocytes, preferably wherein the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is under the control of a cardiomyocyte specific promoter.

25. A composition for the use according to any of claims 21 to 24, wherein the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is a small hairpin RNA (shRNA), small interfering RNA (siRNA), microRNA (miRNA), an antisense oligonucleotide (ASO), an RNA-guided endonuclease system or morpholino oligonucleotide.

26. A composition for the use according to any of claims 21 to 25, wherein the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is a shRNA having a sequence complementary to a portion of SEQ ID NO:

2.

27. A composition for the use according to any of claims 21 to 26, wherein the nucleic acid that is configured to target Runx1 or that specifically binds to a nucleic acid encoding Runx1 is comprised in a vector, preferably a viral vector, more preferably an Adeno-associated virus (AAV) vector, even more preferably an AAV9 viral vector.

28. A composition for the use according to any of claims 11 to 27, wherein the composition comprises one or more pharmaceutically acceptable carriers or excipients.

29. A method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), wherein the method comprises administering to a patient having or at risk of developing HFpEF a therapeutically effective amount of a compound according to the formula:, wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof.

30. A method according to claim 29, wherein the compound is:wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3).

31. A method according to claim 29 or claim 30, wherein the compound is:wherein X is N(R,R1) or O R and R1are each independently C1-C4 alkylor wherein one of R and R1 is hydrogen, preferably wherein N(R,R1) is NH(CH3) 32. A method according to any of claims 29 to 31, wherein the compound is:

33. A method according to any of claims 29 to 32, wherein the compound is Ro5-3335 or Ro24-7429, or a combination thereof.

34. A method according to any of claims 29 to 33, wherein the compound is administered as part of a pharmaceutical composition.

35. A compound for use in a method of prophylaxis or treatment of heart failure with preserved ejection fraction (HFpEF), wherein the method comprises administering to a patient having or at risk of developing HFpEF a therapeutically effective amount of the compound, wherein the compound is:, wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, orN(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3), Y and Z are H, NO2, N(R2,R3), halogen, CF3, OH, CN or lower alkyl or alkoxy, and A, B and C are H, CH3, C2H5, F or CI, and pharmaceutically acceptable salts thereof.

36. A compound for the use according to claim 35, wherein the compound is:wherein X is N(R,R1), O, OR4or SR5, R and R1are each independently N(R2,R3) or C1-C4 alkyl or substituted lower alkyl or an optionally substituted C3-C7-cycloalkyl, aryl, aralkyl or C2-C6-aliphatic hydrocarbon group, this aliphatic group having one or more double or triple bond, or wherein one of R and R1 is hydrogen, or N(R,R1) is an optionally substituted heterocycle with one or more heteroatoms, R2and R3are H, lower alkyl, aryl or aralkyl, R4is lower alkyl or has one of the meanings of R or R1with the exception of H and of N- (R2,R3), R5is H or lower alkyl or has one of the meanings of R or R1with the exception of N- (R2,R3).

37. A compound for the use according to claim 35 or claim 36, wherein the compound is:wherein X is N(R,R1) or O R and R1are each independently C1-C4 alkyl or wherein one of R and R1 is hydrogen, preferably wherein N(R,R1) is NH(CH3) 38. A compound for the use according to any of claims 35 to 37, wherein the compound is:

39. A compound for the use according to any of claims 35 to 38, wherein the compound is Ro5-3335 or Ro24-7429, or a combination thereof.

40. A compound for the use according to any of claims 35 to 39, wherein the compound is administered as part of a pharmaceutical composition.

41. The method, compound for use or composition for use according to any preceding claim, wherein the patient is a human patient, preferably an adult human patient.

42. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the left ventricular ejection fraction (LVEF) of the patient is superior or equal to 40%, optionally to 50%, preferably to 55%, preferably to 60%, even more preferably to 65%, most preferably to 70%.

43. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the patient displays evidence ofcardiac dysfunction characteristic of HFpEF, such as abnormal left ventricular filling and / or elevated filling pressures.

44. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the E / A ratio of the patient is >2.

5.

45. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the patient has an End Diastolic Pressure Volume Relationship (EDPVR) of >12 mmHg.

46. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the patient has an H2FPEF score of |6 or an HFA-PEFF o_kna kb |5, preferably wherein the patient has an H2FPEF score of 6-9 or an HFA-PEFF score of 5-6.

47. The method, compound for use or composition for use according to any preceding claim, wherein prior to administration of the agent, compound or composition the patient has a rest pulmonary _]lehh]nu sa`ca lnaooqna (OBVO) |15 ii Gc kn atan_eoa OBVO |25 ii Gc.

48. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in an improvement in, restoration of, or inhibition of deterioration in diastolic function or cardiac compliance.

49. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in an improvement in diastolic function or cardiac compliance compared to a control value.

50. The method, compound for use or composition for use according to claim 48, wherein the improvement in diastolic function or cadiac compliance is an improvement compared with a placebo.

51. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in a complete restoration of diastolic function or cardiac compliance to values not characteristic of HFpEF, preferably wherein diastolic function or cardiac compliance is measured by E to A wave ratio from M-mode echocardiography and / or End Diastolic Pressure Volume Relationship (EDPVR).

52. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in an E / A ratio of 0.5-2.5, preferably 0.75~2.0, more preferably 0.75~1.5.

53. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in an End Diastolic Pressure Volume Relationship (EDPVR) of <12 mm Hg.

54. The method, compound for use or composition for use according to any preceding claim, wherein administration of the agent, compound or composition results in an improvement in, or resolution or prevention of pulmonary oedema.

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