Molecules targeting LTBP2 and uses thereof

Abstract

The present invention refers to molecules targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2). Further the invention relates to molecules targeting LTBP2 for treatment and diagnosis of diseases associated with LTBP2 expressing fibroblasts. In particular, the invention refers to a nucleic acid encoding molecule targeting LTBP2 and a cell expressing the molecule targeting LTBP2.

Description

[0001] S13211WO / Universität Heidelberg Molecules Targeting LTBP2 and Uses Thereof FIELD OF THE INVENTION The present invention refers to molecules targeting membrane associated latent-transforminggrowth factor beta-binding protein 2 (LTBP2). Further the invention relates to moleculestargeting LTBP2 for treatment and diagnosis of diseases associated with LTBP2 expressing fibroblasts. In particular, the invention refers to a nucleic acid encoding molecule targeting LTBP2 and a cell expressing the molecule targeting LTBP2.Background of the inventionPathological fibrosis is an excessive production of connective tissue in the form of extracellular matrix by fibroblasts and represents a central pathomechanism in various acute or chronic organ diseases. In addition to kidney, lung and liver fibrosis, maladaptive fibrosis in the context of cardiovascular diseases is responsible for one of the most frequent causes of mortality andmorbidity worldwide. This includes various forms of chronic heart failure and tissue remodelingprocesses following myocardial infarction. Maladaptive fibrosis cannot currently be adequately treated since current therapy cannot improve the high mortality and morbidity of cardiac fibrosis since there are only prophylactic measures available (e.g. reduction of cardiac risk factors). Thus, there is a need for treatment of fibrosis and myocardial infarction. Objectives and Summary of the Invention The present invention solves the above-mentioned problem by providing new methods for diagnosis and treatment of diseases associated with LTBP2 expressing cells, in particular, LTBP2 expressing fibroblasts. Accordingly, the invention relates to molecules targeting LTBP2for use in treatment or prevention of fibrosis, chronic heart failure, and / or myocardial infarction.LTBP2 is a very cell- and disease-specific protein that is a component of central fibrosis signaling pathways, it was not considered so far for treatment methods directly interfering with activated fibroblasts, since the protein is secreted from the activated fibroblasts and it was so far assumed that after secretion it is freely released into the extracellular spaceand not further associated with the activated fibroblasts.Since LTBP2 is hardly present in healthy tissue, it can be expected that LTBP2-directed treatment strategies show no or only minimal side effects. The use of LTBP2 as a molecular or LTBP2-expressing activated fibroblasts as cellular therapy target enables a specific regulation ofthe production of extracellular matrix and thus regulation of fibrosis.LTBP2 is secreted by the activated fibroblasts. However, so far, it was not known that the secreted protein remains associated with the membrane of the activated fibroblasts. Contrary to S13211WO / Universität Heidelberg what is described in literature the inventors showed for the first time that LTBP2 is closely associated with the plasma membrane which makes it a suitable target for immune therapy. Thus, the inventors for the first time show that LTBP2, due to its membrane association in activated fibroblast can be targeted by diagnostic and treatment approaches which exploittargeting activated fibroblasts marked by membrane associated LTBP2. In particular, binding tomembrane associated LTBP2 allows regulating and / or destroying directly cells which exhibit membrane associated LTBP2. As an example, the inventors could successfully show that an anti-LTBP2 CAR T cell can be used for killing activated human fibroblasts.Thus, a first aspect of the invention refers to the use of a molecule targeting membraneassociated LTBP2 in treatment of a disease associated with LTBP2 expressing fibroblasts. In some embodiments, LTBP2 may be associated to the membrane in the LTBP2 expressing fibroblasts. In some embodiments, molecule targeting membrane associated LTBP2 may be selected from the group consisting of a chimeric antigen receptor (CAR), an antibody, a bispecific antibody, a bispecific antibody linking T cells with target cells (bispecific T cell engager;BiTE), a single domain antibody (sdAb, nanobody) antibody-drug conjugate (ADC) and a small molecule.Moreover, these molecules may recognize and bind to the membrane-associated LTBP2,thereby initiating a therapeutic response. In some embodiments, the molecule targeting membrane associated LTBP2 may comprise a single-chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence ofSEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6.In some embodiments, the molecule targeting membrane associated LTBP2 may comprise asingle-chain variable fragment (scFv) comprising a variable heavy chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 7. In some embodiments, the molecule targeting membrane associated LTBP2 may comprise asingle-chain variable fragment (scFv) comprising a variable light chain having a sequenceidentical to or at least 80% identical to the sequence set out in SEQ ID NO: 8. In some embodiments, the molecule targeting membrane associated LTBP2 may be a CAR comprising from N-terminus to C-terminus, the scFv, optionally at least one co-stimulatorydomain and an activating domain, wherein the scFv may comprise a variable heavy chaincomprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid S13211WO / Universität Heidelberg sequence of SEQ ID NO: 6, and / or a variable heavy chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 7, and / or a variable light chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 8. In some preferred embodiments, at least one co-stimulatory domain may be selected from thegroup consisting of CD28 and 4-1BB. In some preferred embodiments, the activating domainmay be CD3zeta. In some embodiments, the molecule targeting membrane associated LTBP2 may be a CAR comprising from N-terminus to C-terminus, the scFv domain as described herein, a CD28 co-stimulatory domain, a 4-1BB co-stimulatory domain and a CD3zeta activating domain, whereinthe scFv may comprise a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO: 6, or a variable heavy chain havinga sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 7, or a variable light chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 8. In some embodiments, CD28 co-stimulatory domain may have a sequence which may be at least 80% identical to SEQ ID NO.: 11. In some embodiments, the 4-1BB co-stimulatory domain may have a sequence which may be at least 80% identical to SEQ ID NO.:12. In some embodiments, the CD3zeta activating domain may have a sequence which may be at least 80% identical to SEQ ID NO.: 13. In some embodiments, the disease associated with LTBP2 expressing fibroblasts may befibrosis.In some embodiments, the disease associated with LTBP2 expressing fibroblasts may be selected from the group consisting of fibrosis in the context of the cardiovascular disease, kidney fibrosis, lung fibrosis and liver fibrosis. Fibrosis in the context of the cardiovascular disease may be selected from the group consisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiac fibrosis.The molecule targeting membrane associated LTBP2 may recognize and bind to the membrane-associated LTBP2 and thereby initiating a therapeutic response. For example, at least one echocardiographic parameter may be improved, typically after myocardial infarction. In one embodiment, the echocardiographic parameter is selected from the group consisting ofejection fraction, fractional area change and end-diastolic volume. In one embodiment, theejection fraction is left ventricular ejection fraction and / or right ventricular ejection fraction. In one embodiment, the end-diastolic volume is left ventricular end-diastolic volume and / or right ventricular end-diastolic volume. S13211WO / Universität Heidelberg In some specific embodiments, CAR targeting LTBP2 may comprise from N-terminus to C- terminus, the scFv, at least one co-stimulatory domain, and an activating domain, wherein the scFv may comprise a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 havingthe amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acidsequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6, or a variable heavy chain having a sequence identical to the sequence or at least 80% identical to the sequence set out in SEQ ID NO: 7, or a variable light chain having a sequence identical to the sequence or at least 80%identical to the sequence set out in SEQ ID NO: 8.In some specific embodiments, at least one co-stimulatory domain in any of CAR targeting LTBP2 may be selected from the group consisting of CD28 and 4-1BB. In some specific embodiments, the activating domain in any of CAR targeting LTBP2 may be CD3zeta.In specific examples, the molecule targeting membrane associated LTBP2 may be a CARcomprising from N-terminus to C-terminus, the scFv, CD28 co-stimulatory domain, 4-1BB co- stimulatory domain, and CD3zeta activating domain, wherein the scFv may comprise a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence ofSAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO:4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6, or a variable heavy chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 7, or a variable light chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 8.In certain embodiments, the CD28 co-stimulatory domain in any of the CAR targeting LTBP2 canhave a sequence which may be at least 80% identical to SEQ ID NO.: 11. In some embodiments, the 4-1BB co-stimulatory domain in any of the CAR targeting LTBP2 can have a sequence which may be at least 80% identical to SEQ ID NO.: 12. In some embodiments, the CD3zeta activating domain in any of the CAR targeting LTBP2 canhave a sequence which may be at least 80% identical to SEQ ID NO.: 13.In another aspect, a nucleic acid may encode any of the CARs disclosed herein. Other aspects of the present disclosure provide modified cells expressing the CAR described herein. In some embodiments, the cell can be a T cell, a NK cell, a dendritic cell, a macrophage, a monocyte, a B cell, a neutrophil, an eosinophil, a basophil, a mast cell, a myeloid derivedsuppressor cell, a mesenchymal stem cell, a precursor thereof, or a combination.Another aspect of the invention refers to the use of molecule targeting membrane associated LTBP2 for treatment and diagnosis of diseases associated with LTBP2 expressing fibroblasts. S13211WO / Universität Heidelberg Figure Legends Figure 1: LTBP2 mRNA and protein expression is strongly induced in the fibrotic zone following myocardial infarction A) Time course of LTBP2 mRNA expression in infarct vs. borderzone showing a peak level inmurine hearts between day 7 and 14 post myocardial infarction. ns= not significant, **p<0.01,***p<0.001 B) Quantification of the dynamic protein LTBP2 expression in murine scar-areas following ischemia / reperfusion injury (I / R, transient LAD-occlusion) by fluorescence intensity, *p<0.05, ***p<0.001. C-D) Representative immunofluorescence stainings of different time points after permanent (C) or transient (D) LAD occlusion as murine models for myocardial infarction,the two left-sided rows of each panel represent whole section overviews (scale bar = 1mm), theright-sided detailed magnifications (scale bar = 100µm). WGA = Wheat germ agglutinin. Figure 2: LTBP2 is very specific for activated fibroblasts and nearly solely expressed in diseased hearts.Analysis of CITE-Seq protein expression data sets [1] from human cardiac tissue. A) UMAP Plotindicating LTBP2 expression in various cardiac cell types. B) LTBP2 expression within fibroblast subclusters showing an overlap with activated fibroblast states [1]. C) Quantification of total LTBP2 protein expression in human healthy vs. acute myocardial infarction, chronic ischemic and non-ischemic cardiomyopathy.Figure 3: LTBP2 is expressed in association with the plasma membrane of humanfibroblasts in vitro following TGFβ-stimulation. A) Experimental design of in vitro experiments using primary, human ventricular fibroblasts. B) Representative confocal microscopy images of an unstimulated and TGFβ-stimulated fibroblast showing LTBP2 at the plasma membrane. C) Quantification of LTBP2 protein expression inhuman fibroblasts after TGFβ-stimulation in different concentrations. D) Expression of LTBP2determined by flow cytometry of non-permeabilized cells with and without previous growth-factor stimulation in activated fibroblasts. Figure 4: Generation and successful application of specific anti-LTBP2 CAR-T cells in vitro and in an in vivo mouse model of myocardial infarction.A) A schematic overview of CAR targeting LTBP2. B) Experimental results of the successful,dose-dependent killing of activated human fibroblasts in vitro. Mock = control treated T cells. Right panel shows killing properties normalized to mock T cells. C) In vivo experimental results of murine anti-LTBP2 CAR T cells compared to PBS control in mice after minimal-invasive LAD occlusion [2] as a model for myocardial infarction (MI). CAR-T or PBS injection were performedat day 6 following MI. Data represent cardiac function assessed by echocardiography at day 31post MI. FAC (SAX) = fractional area change in parasternal short axis views; LVEDV = left ventricular end-diastolic volume; LVEF = left ventricular ejection fraction, PLAX = parasternal long axis views. D) Analysis of the aortic sinus diameter and ascending aorta diameter (Aorta S13211WO / Universität Heidelberg asc. diam.) showed no adverse effects, such as dilation, on the large vessels within the same cohort as described in C. Figure 5: A) Neutralizing anti-LTBP2-anti-hCD3ε antibody: Left panel showing schematic of the antibodydesign and construction of the LTBP2-neutralising anti-LTBP2 x anti-hCD3ε bispecific bivalentantibody (anti-human). Right panel demonstrates the mode of action for T-cell mediated killing of LTBP2-expressing fibroblasts. Fab = fragment antigen-binding region; Fc = fragment crystallizable region; KiHss = knobs-into-holes binding; scFv = single chain variable fragment. B) anti-LTBP2-anti-hCD3ε tetravalent antibody: Left panel showing schematic of the tetravalentantibody design and construction of the anti-LTBP2 x anti-hCD3ε bispecific tetravalent antibody(anti-human). Right panel demonstrates the mode of action for T-cell mediated killing of LTBP2- expressing fibroblasts. scFv = single chain variable fragment. C) anti-LTBP2 NK-cell engaging antibody: Left panel showing schematic of the antibody design and construction of the anti-LTBP2 bifunctional NK-cell engaging antibody (anti-human). Rightpanel demonstrates the mode of action for CD16 mediated killing of LTBP2-expressingfibroblasts. Fc = fragment crystallizable region; hIgG1 = human immunoglobulin 1; NK = natural killer; scFv = single chain variable fragment. Figure 6: Functional evaluation of human bispecific antibodies targeting LTBP2 in vitroand in an “ex vivo” living myocardial slice modelA) Schematic of the experimental design for killing assays using the xCELLigence platform and respective gold microelectrode plates for continuous impendence detection. Human primary fibroblasts and heterologous healthy human T-cells and NK cells were used. B-D) Results of human killing assays for the three different antibody constructs. Number if the construct indicatedin legends including the respective concentration (nM). Cell-index values were normalized tofibroblast-only replicates to calculate relative viability (%). The black arrow indicates the addition of bsMAbs, after the T- and NK cells were pipetted into the wells. Dashed lines represent control conditions (fibroblasts + T- or NK cells). All groups and concentrations were performed in n=6 replicates and means were calculated for the real time values. E) Experimental design for human“ex-vivo” living myocardial slice model. Homologous T- and NK cells were isolated prior to surgeryfor every patient individually (n=4). bsMAb = bispecific monoclonal antibody construct; LVAD = left ventricular assist device; NK = natural killer cell. F) Representative images of a living myocardial slice clamped onto an elastic frame in the direction of the myocardial fibers and placed into a 12-well plate under cell-culture conditions for incubation with T-cells and the anti-CD3 / anti-LTBP2 bsMAb (No.: 15452.1). G) Immunofluorescence staining of the slice indicating apoptosisby cleaved caspase 3 signal (Casp3, light grey) and DAPI (dark grey) for nuclei detection. The white asterisks indicates a co-signal of DAPI and Casp3 defining a granulated nuclei of an apoptotic cell within the fibrotic heart failure tissue. H) Quantification of the immunofluorescence stainings (see G) by scanning the whole slice and semi-automated analysis using QuPathsoftware. N = 1-6 slices / patient. S13211WO / Universität Heidelberg Figure 7: A) anti-CD3 / anti-LTBP2 tetravalent antibody: Left panel showing schematic of the tetravalent antibody design and construction. Right panel demonstrates the mode of action for T-cell mediated killing of LTBP2-expressing fibroblasts. scFv = single chain variable fragment.B) anti-CD16 / anti-LTBP2 NK-cell engaging antibody: Left panel showing schematic of theantibody design and construction. Right panel demonstrates the mode of action for CD16 mediated killing of LTBP2-expressing fibroblasts. Fc = fragment crystallizable region; mIgG2a = murine immunoglobulin 2a; NK = natural killer; scFv = single chain variable fragment. C) costimulatory anti-LTBP2 anti-CD28 antibody: Left panel showing schematic of the antibodydesign and construction. Right panel demonstrates the mode of action for T-cell mediated killingof LTBP2-expressing fibroblasts. scFv= single chain variable fragment. Figure 8: Functional and therapeutic evaluation of murine bispecific antibodies targeting LTBP2A-B) Results of murine killing assays using xCELLigence assays and primary murine cardiacfibroblasts from the same mice (C57BL / 6N) as the used T- and NK cells isolated from splenocytes. Viability was calculated from cell-index values after normalization to fibroblast-only control wells. The black arrow indicates the timepoint of the T / NK-cell and bsMAb application. Numbers of the respective constructs are stated in legends including the used concentrations (nM). C)Experimental design of the murine in vivo study. Echo = echocardiography; mi-IR = minimal-invasive ischemia / reperfusion; TnT = cardiac troponin T. D) Time course of left ventricular ejection fraction (LVEF) for all groups at the indicated timepoints in repeated measurements. E-K) Parameters from day 28. Cardiac function was quantified from left-ventricular parasternal long- axis B-mode traces. EF = ejection fraction; HW / TL = heart weight normalized to tibia length; V,d= end-diastolic volume of the left ventricle; V,s = end-systolic volume of the left ventricle; Area,d= end-diastolic area of the left ventricle. N=6 / group, no animal was excluded from final analysis, statistical analysis performed using one-way ANOVA with Tukey's multiple comparisons test. P- values <0.05 shown above graphs.Figure 9: LTBP2 knock-out mice show improved infarct healing and attenuated cardiacfibrosis after myocardial ischemia-reperfusion injury A) Experimental design: LTBP2-KO and WT mice (C57BL / 6J) underwent 60 minutes of echocardiography-guided ischemia-reperfusion injury (IR) to induce myocardial infarction. Cardiac function was assessed via echocardiography on days 1, 7, 14, and 28 post-IR. Mice weresacrificed on day 28 for histological analysis. B) Representative echocardiographic images of theLV from WT and LTBP2-KO mice at day 1 and day 28 post-IR. C) Infarct size at day 1 and 28 determined by echocardiography. D) Quantification of key echocardiographic parameters (ejection fraction, global longitudinal strain, and end-systolic volume) is depicted. E) Representative images of different planes throughout the LV in WT and KO hearts using a Picrosirius-Red staining.Fibrotic areas across the 4 different planes of the LV, expressed as a percentage of the LV wall,were averaged for each mouse and quantified in (F). G) Representative images of POSTN immunofluorescence staining in WT and KO hearts at day 28 post-IR. H) Quantification of POSTN S13211WO / Universität Heidelberg positive area in percentage of the LV wall. Statistics: n = 10 per group; statistical analysis by one- way ANOVA with Tukey’s multiple comparisons test; p < 0.05. KO = genetic knock out; LV = left ventricle; WT = wildtype.Figure 10: Reduced Liver Fibrosis in LTBP2 KO mice in a murine model of liver cirrhosisand fibrosis A) Experimental Setup: Liver injury was induced in 14- to 16-week-old male and female LTBP2- KO and WT mice via intraperitoneal injections of CCl₄, administered three times per week for four weeks (2.5 µl / g body weight; 25% CCl₄ in corn oil). B) Graphs showing the body weight courseover time. C-D) Serum liver enzyme levels (AST and ALT) at day 28 measured in IU / l. E)Representative Picrosirius Red-stained liver sections at day 28 illustrating the amount of fibrosis (dark grey), which is further quantified in (F). G) Representative images showing immunofluorescence staining for DAPI (upper row) and CD68 (lower row). H) CD68 positive cells were annotated, counted and quantified in H). Statistics: n = 8 per group; statistical analysis byone-way ANOVA with Tukey’s multiple comparisons test; P-values shown above columns, p <0.05 defined as significant. AST = aspartate transaminase; ALT = alanine transaminase; CCl₄ = carbon tetrachloride; KO = genetic knock out; WT = wildtype. Figure 11: A-G: Exemplary multifunctional antibody constructs. DETAILED DESCRIPTION OF THE INVENTION Definitions Before the invention is described in detail with respect to some of its preferred embodiments, thefollowing general definitions are provided.The present invention as illustratively described in the following may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. The present invention will be described with respect to particular embodiments and with reference to certain figures but the invention is not limited thereto but only by the claims.Where the term “comprising” is used in the present description and claims, it does not excludeother elements. For the purposes of the present invention, the term “consisting of” is considered to be a preferred embodiment of the term “comprising of”. If hereinafter a group is defined to comprise at least a certain number of embodiments, this is also to be understood to disclose a group which preferably consists only of these embodiments.For the purposes of the present invention, the term “obtained” is considered to be a preferredembodiment of the term “obtainable”. If hereinafter e.g. a compound is defined to be obtainable from a specific source, this is also to be understood to disclose a compound which is obtained from this source. S13211WO / Universität Heidelberg Further, as used in the following, the terms "preferably", "more preferably", "most preferably", "particularly", "more particularly", "specifically", "more specifically" or similar terms are used in conjunction with optional features, without restricting further possibilities. Thus, features introduced by these terms are optional features and are not intended to restrict the scope of theclaims in any way. The invention may, as the skilled person will recognize, be performed byusing alternative features. Where an indefinite or definite article is used when referring to a singular noun, e.g. “a”, “an” or “the”, this includes a plural of that noun unless something else is specifically stated. The terms “about” or “approximately” in the context of the present invention denote an interval of accuracythat the person skilled in the art will understand to still ensure the technical effect of the featurein question. The term typically indicates deviation from the indicated numerical value of ±10%, and preferably of ±5%. Technical terms are used by their common sense. If a specific meaning is conveyed to certain terms, definitions of terms will be given in the following in the context of which the terms areused.“Sequence identity” or “percentage identity” and “sequence similarity” can be determined by alignment of two peptide or two nucleotide sequences using global or local alignment algorithms. For example, sequence similarity or identity can be determined by searching against databases such as FASTA, BLAST, etc., but hits should be retrieved and aligned pairwise to comparesequence identity. Two proteins or two protein domains, or two nucleic acid sequences thatdescribed herein as at least 80% identical to a reference sequence, include peptide sequences and nucleic sequences which are at least 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% preferably at least 90%, 95%, 98%, 99% identical to the reference sequence. For the Molecules Targeting LTBP2 described herein the named antigen binding molecule includes any of the amino sequences set our herein, or variants or homologs that maintain the activity (e.g., within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the binding molecule set out herein). In some embodiments, variants or homologshave at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity acrossthe whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to the amino acid sequence set out herein.“Nucleic acid" generally means a polymer of DNA or RNA, which can be single-stranded ordouble-stranded, synthesized or obtained (e.g., isolated and / or purified) from natural sources, which can contain natural, non-natural or altered nucleotides, and which can contain a natural, non-natural or altered internucleotide linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of anunmodified oligonucleotide. Preferably, the nucleic acids described herein are recombinant. Asused herein, the term "recombinant" refers to (i) molecules that are constructed outside living cells by joining natural or synthetic nucleic acid segments to nucleic acid molecules that can S13211WO / Universität Heidelberg replicate in a living cell, or (ii) molecules that result from the replication of those described in (i) above. For purposes herein, the replication can be in vitro replication or in vivo replication. The nucleic acids can be constructed based on chemical synthesis and / or enzymatic ligation reactions using procedures known in the art or commercially available (e.g. from Genscript,Thermo Fisher and similar companies). See, for example Sambrook et al., a nucleic acid can bechemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed upon hybridization (e.g., phosphorothioate derivatives and acridine substituted nucleotides). In some embodiments, the amino acid sequence of the molecules described herein may comprise one or more phenotypically silent substitutions. “Phenotypically silent substitutions” are also named “conservative amino acid substitutions". The concept of "conservative amino acid substitutions" is understood by the skilled artisan, and preferably means that codons encodingpositively-charged residues (H, K, and R) are substituted with codons encoding positively-charged residues, codons encoding negatively- charged residues (D and E) are substituted with codons encoding negatively-charged residues, codons encoding neutral polar residues (C, G, N, Q, S, T, and Y) are substituted with codons encoding neutral polar residues, and codons encoding neutral non-polar residues (A, F, I, L, M, P, V, and W) are substituted with codonsencoding neutral non-polar residues. These variations can spontaneously occur, be introducedby random mutagenesis, or can be introduced by directed mutagenesis. Those changes can be made without destroying the essential characteristics of these antigen binding molecules described herein, i.e. the binding functionality. The ordinarily skilled artisan can readily and routinely screen variant amino acids and / or the nucleic acids encoding them to determine ifthese variations substantially reduce or destroy the binding capacity by methods known in theart. The skilled person understands, that also the nucleic acid encoding the antigen binding molecules described herein may be modified. Useful modifications in the overall nucleic acid sequence include codon optimization of the sequence. Alterations may be made which lead to conservative substitutions within the expressed amino acid sequence. Usually, additions anddeletions should not be performed in the CDR regions."Single-chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain and in either orientation {e.g., VL-VH or VH-VL). Generally, the scFv polypeptide further comprises a polypeptide linkerbetween the VH and VL domains which enables the scFv to form the desired structure forantigen binding. A “Fab” also termed “Fab fragment” or “Fab region” consists of the VL, VH, CL and CH1 domains. An F(ab')2 fragment comprises two Fab fragments linked by a disulfide bridge at thehinge region.“Engineered” refers to a modification of a part of a molecule, such as a domain. In particular, it refers to protein sequence that has its amino acid sequence modified actively, non-naturally, in order to improve desired biochemical characteristics. S13211WO / Universität Heidelberg “Multispecific antigen binding molecule” is an antigen binding molecule having at least two different antigen binding sites, each site with a different binding specificity.“Bispecific antigen binding molecule” is an antigen binding molecule having at two differentantigen binding sites, each site with a different binding specificity. “Bispecific antibody” is an antibody having two different antigen binding sites, each site with a different binding specificity. A bispecific antibody can be a full-length antibody or an antibodyfragment, and the different binding sites may bind each to a different antigen or the differentbinding sites may bind to two different epitopes of the same antigen. In a preferred embodiment, the different binding sites bind each to a different antigen, e.g. to a LTBP2 and to an immune cell protein, such as CD3, CD28 or CD16.“Multispecific antibody” is an antibody having at least two different antigen binding sites, eachsite with a different binding specificity. A multispecific antibody can be a full-length antibody or an antibody fragment, and the different binding sites may bind each to a different antigen or the different binding sites may bind to two different epitopes of the same antigen.The term “antibody-drug conjugate” or "ADC" as used herein refers to an antibody or antibodyfragment to which toxins (or drugs) have been linked. In an ADC, toxins are conjugated to the antibody or antibody fragment by cleavable or non-cleavable linkers. Conjugation of the linker to the antibody may for example rely on the presence of lysine and cysteine residues within the polypeptide structure of the antibody as the point of conjugation. Reactive groups on the linkercan e.g. be conjugated to the side chain of lysine residues through amide or amidine bondformation. Conjugation via cysteine residues requires a partial reduction of the antibody. Alternatively, site-specific enzymatic conjugation can be used. This requires enzymes that react with the antibody and can induce site- or amino acid sequence-specific modifications. Peptide sequences recognized by these enzymes may have to be inserted into the geneticallyengineered antibodies or fragments to be conjugated. Enzymes which have been used for suchpurpose are sortase, transglutaminase, galactosyltransferase, sialyltransferase and tubulin- tyrosine ligase. An overview of ADC linker conjugation and toxins can be found in Ponziani et al, 2020 (Ponziani et al.2020). An overview of conjugation of toxins to antibody fragments can be found in Aguiar et al, 2018 (Aguiar et al.2018). An individual or subject treated in accordance with this invention refers to vertebrates, particularly members of a mammalian species, and includes but is not limited to domestic animals, sports animals, and primates, including humans. In one embodiment, the subject treated in accordance with this invention is a mammal. In one embodiment, the subject treated inaccordance with this invention is a human. In another embodiment, the subject treated inaccordance with this invention is a non-human mammal. On aspect refers to a molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) for use as a medicament. S13211WO / Universität Heidelberg One aspect of the invention relates to a molecule targeting membrane associated latent- transforming growth factor beta-binding protein 2 (LTBP2) for use in treatment of a disease associated with LTBP2 expressing fibroblasts.Another aspect of the disclosure relates to the molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2). The term “latent-transforming growth factor beta-binding protein 2”, abbreviated LTBP2, refers to human UniProt Q14767 or any mammalian equivalent, such as mouse UniProt O08999: Human (SEQ ID NO.: 27):>sp|Q14767|LTBP2_HUMAN Latent-transforming growth factor beta-binding protein2 OS=Homo sapiens OX=9606 GN=LTBP2 PE=1 SV=3 MRPRTKARSPGRALRNPWRGFLPLTLALFVGAGHAQRDPVGRYEPAGGDANRLRRPGGSY PAAAAAKVYSLFREQDAPVAGLQPVERAQPGWGSPRRPTEAEARRPSRAQQSRRVQPPAQ TRRSTPLGQQQPAPRTRAAPALPRLGTPQRSGAAPPTPPRGRLTGRNVCGGQCCPGWTTANSTNHCIKPVCEPPCQNRGSCSRPQLCVCRSGFRGARCEEVIPDEEFDPQNSRLAPRRWAERSPNLRRSSAAGEGTLARAQPPAPQSPPAPQSPPAGTLSGLSQTHPSQQHVGLSRTVRL HPTATASSQLSSNALPPGPGLEQRDGTQQAVPLEHPSSPWGLNLTEKIKKIKIVFTPTIC KQTCARGHCANSCERGDTTTLYSQGGHGHDPKSGFRIYFCQIPCLNGGRCIGRDECWCPA NSTGKFCHLPIPQPDREPPGRGSRPRALLEAPLKQSTFTLPLSNQLASVNPSLVKVHIHHPPEASVQIHQVAQVRGGVEEALVENSVETRPPPWLPASPGHSLWDSNNIPARSGEPPRPLPPAAPRPRGLLGRCYLNTVNGQCANPLLELTTQEDCCGSVGAFWGVTLCAPCPPRPASPV IENGQLECPQGYKRLNLTHCQDINECLTLGLCKDAECVNTRGSYLCTCRPGLMLDPSRSR CVSDKAISMLQGLCYRSLGPGTCTLPLAQRITKQICCCSRVGKAWGSECEKCPLPGTEAF REICPAGHGYTYASSDIRLSMRKAEEEELARPPREQGQRSSGALPGPAERQPLRVVTDTWLEAGTIPDKGDSQAGQVTTSVTHAPAWVTGNATTPPMPEQGIAEIQEEQVTPSTDVLVTLSTPGIDRCAAGATNVCGPGTCVNLPDGYRCVCSPGYQLHPSQAYCTDDNECLRDPCKGKG RCINRVGSYSCFCYPGYTLATSGATQECQDINECEQPGVCSGGQCTNTEGSYHCECDQGY IMVRKGHCQDINECRHPGTCPDGRCVNSPGSYTCLACEEGYRGQSGSCVDVNECLTPGVC AHGKCTNLEGSFRCSCEQGYEVTSDEKGCQDVDECASRASCPTGLCLNTEGSFACSACENGYWVNEDGTACEDLDECAFPGVCPSGVCTNTAGSFSCKDCDGGYRPSPLGDSCEDVDECEDPQSSCLGGECKNTVGSYQCLCPQGFQLANGTVCEDVNECMGEEHCAPHGECLNSHGSFF CLCAPGFVSAEGGTSCQDVDECATTDPCVGGHCVNTEGSFNCLCETGFQPSPESGECVDI DECEDYGDPVCGTWKCENSPGSYRCVLGCQPGFHMAPNGDCIDIDECANDTMCGSHGFCD NTDGSFRCLCDQGFEISPSGWDCVDVNECELMLAVCGAALCENVEGSFLCLCASDLEEYDAQEGHCRPRGAGGQSMSEAPTGDHAPAPTRMDCYSGQKGHAPCSSVLGRNTTQAECCCTQGASWGDACDLCPSEDSAEFSEICPSGKGYIPVEGAWTFGQTMYTDADECVIFGPGLCPNG RCLNTVPGYVCLCNPGFHYDASHKKCEDHDECQDLACENGECVNTEGSFHCFCSPPLTLD LSQQRCMNSTSSTEDLPDHDIHMDICWKKVTNDVCSEPLRGHRTTYTECCCQDGEAWSQQ CALCPPRSSEVYAQLCNVARIEAEREAGVHFRPGYEYGPGPDDLHYSIYGPDGAPFYNYLGPEDTVPEPAFPNTAGHSADRTPILESPLQPSELQPHYVASHPEPPAGFEGLQAEECGILNGCENGRCVRVREGYTCDCFEGFQLDAAHMACVDVNECDDLNGPAVLCVHGYCENTEGSY RCHCSPGYVAEAGPPHCTAKE Mouse (SEQ ID NO.: 28):>sp|O08999|LTBP2_MOUSE Latent-transforming growth factor beta-binding protein2 OS=Mus musculus OX=10090 GN=Ltbp2 PE=1 SV=2 S13211WO / Universität Heidelberg MRAPTTARCSGCIQRVRWRGFLPLVLAVLMGTSHAQRDSIGRYEPASRDANRLWHPVGSH PAAAAAKVYSLFREPDAPVPGLSPSEWNQPAQGNPGWLAEAEARRPPRTQQLRRVQPPVQ TRRSHPRGQQQIAARAAPSVARLETPQRPAAARRGRLTGRNVCGGQCCPGWTTSNSTNHC IKPVCQPPCQNRGSCSRPQVCICRSGFRGARCEEVIPEEEFDPQNARPVPRRSVERAPGPHRSSEARGSLVTRIQPLVPPPSPPPSRRLSQPWPLQQHSGPSRTVRRYPATGANGQLMSNALPSGLELRDSSPQAAHVNHLSPPWGLNLTEKIKKIKVVFTPTICKQTCARGRCANSCEK GDTTTLYSQGGHGHDPKSGFRIYFCQIPCLNGGRCIGRDECWCPANSTGKFCHLPVPQPD REPAGRGSRHRTLLEGPLKQSTFTLPLSNQLASVNPSLVKVQIHHPPEASVQIHQVARVR GELDPVLEDNSVETRASHRPHGNLGHSPWASNSIPARAGEAPRPPPVLSRHYGLLGQCYLSTVNGQCANPLGSLTSQEDCCGSVGTFWGVTSCAPCPPRQEGPAFPVIENGQLECPQGYKRLNLSHCQDINECLTLGLCKDSECVNTRGSYLCTCRPGLMLDPSRSRCVSDKAVSMQQGL CYRSLGSGTCTLPLVHRITKQICCCSRVGKAWGSTCEQCPLPGTEAFREICPAGHGYTYS SSDIRLSMRKAEEEELASPLREQTEQSTAPPPGQAERQPLRAATATWIEAETLPDKGDSR AVQITTSAPHLPARVPGDATGRPAPSLPGQGIPESPAEEQVIPSSDVLVTHSPPDFDPCFAGASNICGPGTCVSLPNGYRCVCSPGYQLHPSQDYCTDDNECMRNPCEGRGRCVNSVGSYSCLCYPGYTLVTLGDTQECQDIDECEQPGVCSGGRCSNTEGSYHCECDRGYIMVRKGHCQ DINECRHPGTCPDGRCVNSPGSYTCLACEEGYVGQSGSCVDVNECLTPGICTHGRCINME GSFRCSCEPGYEVTPDKKGCRDVDECASRASCPTGLCLNTEGSFTCSACQSGYWVNEDGT ACEDLDECAFPGVCPTGVCTNTVGSFSCKDCDQGYRPNPLGNRCEDVDECEGPQSSCRGGECKNTEGSYQCLCHQGFQLVNGTMCEDVNECVGEEHCAPHGECLNSLGSFFCLCAPGFASAEGGTRCQDVDECAATDPCPGGHCVNTEGSFSCLCETASFQPSPDSGECLDIDECEDRED PVCGAWRCENSPGSYRCILDCQPGFYVAPNGDCIDIDECANDTVCGNHGFCDNTDGSFRC LCDQGFETSPSGWECVDVNECELMMAVCGDALCENVEGSFLCLCASDLEEYDAEEGHCRP RVAGAQRIPEVRTEDQAPSLIRMECYSEHNGGPPCSQILGQNSTQAECCCTQGARWGKACAPCPSEDSVEFSQLCPSGQGYIPVEGAWTFGQTMYTDADECVLFGPALCQNGRCSNIVPGYICLCNPGYHYDASSRKCQDHNECQDLACENGECVNQEGSFHCLCNPPLTLDLSGQRCVN TTSSTEDFPDHDIHMDICWKKVTNDVCSQPLRGHHTTYTECCCQDGEAWSQQCALCPPRS SEVYAQLCNVARIEAERGAGIHFRPGYEYGPGLDDLPENLYGPDGAPFYNYLGPEDTAPE PPFSNPASQPGDNTPVLEPPLQPSELQPHYLASHSEPPASFEGLQAEECGILNGCENGRCVRVREGYTCDCFEGFQLDAPTLACVDVNECEDLNGPARLCAHGHCENTEGSYRCHCSPGYVAEPGPPHCAAKE In one embodiment, LTBP2 refers to a protein having the amino acid sequence which is identical or at least 70% identical to SEQ ID NO: 27.The term “membrane associated LTBP2” refers to LTBP2 fibroblasts and which is associatedwith the plasma membrane of the fibroblasts. Associated with the membrane means that it is retained at the cell membrane, e.g. of the activated fibroblast, i.e. is accumulating at the extracellular side of the membrane of the activated fibroblast. Typically, membrane associated LTBP2 associates with the membrane after secretion, more particularly activated fibroblastssecrete LTBP2 which then binds or accumulates to the outside surface of the plasma membraneof the activated fibroblasts. “LTBP2 expressing fibroblasts” as used herein refers to fibroblasts which express LTBP2 and which is associated with the membrane of these fibroblasts. The term “disease associated with LTBP2 expressing fibroblasts” as used herein refers to diseases in which LTBP2 is abnormally expressed by fibroblasts. LTBP2 expression is ahallmark of fibrosis. Hence, the term disease associated with LTBP2 expressing fibroblastsincludes fibrosis, including for example cardiovascular fibrosis, perivascular fibrosis, vascular fibrosis, kidney fibrosis, lung fibrosis and liver fibrosis. Since LTBP2 is expressed in the fibroblasts of the blood vessels, the term disease associated with LTBP2 expressing fibroblasts S13211WO / Universität Heidelberg also includes indications such as vascular fibrosis, perivascular fibrosis, atherosclerosis and stroke. In some embodiments, the disease associated with LTBP2 expressing fibroblasts may be fibrosis, defined as any disease not directly induced by cancer cells. Tumour-associated matrixdeposition is induced by cancer cells that drive the differentiation of specific tumour-associatedfibroblasts (also termed “cancer-associated fibroblasts”) in order to promote the malignant microenvironment and spreading of tumour cells. Fibrosis of any other origin is not induced by cancer cells and therefore a biologically different entity. Fibrosis consists of organ diseases mediated or associated with fibrotic deposition ofextracellular matrix other than matrix deposition within malignant tumours (=cancer), regardlessof the respective organ: Fibrosis consists of variable diseases e.g. cardiac fibrosis, kidney fibrosis, lung fibrosis, bone- marrow fibrosis, skin fibrosis (e.g. systemic sclerosis or scar formation) and liver fibrosis. It can occur in any organ and can be induced by e.g. tissue damage (such as infarct, stroke, radiation,heat or burning, iatrogenic like surgery), hypoxia, auto-immunity, inflammation, immune-activation, degeneration, chemical toxicity, drug side effects, or can arise from unknown effects called idiopathic fibrosis. Tumour-tissue matrix deposition is induced by cancer cells and consists of diseases e.g. cardiac tumours, kidney tumours, lung cancer (small cell lung carcinoma, bronchial carcinoma),leukaemia, skin tumours (malignant melanoma, basaliona), and liver tumours (hepatocellularcarcinoma, cholangio cellular carcinoma) and any other malignant organ cancer associated with cancer-associated fibroblasts promoting cancer cell metastasis etc. In some embodiments, the disease associated with LTBP2 expressing fibroblasts may be selected from the group consisting of fibrosis in the context of the cardiovascular disease, kidneyfibrosis (such as nephritis, thrombotic microangiopathy, minimal-change glomerulopathy,membranous glomerulopathy, glomerulosclerosis, diabetic nephropathy, lupus nephritis, amyloidosis, polycystic kidney disease, IgA nephropathy Alport syndrome), lung fibrosis (ideopatic, chronic-obstructive pulmonary disease, acute respiratory distress syndrome, cystic fibrosis, pulmonary hypertension, legionella infections, tuberculosis, drug-induced fibrosis, etc.),liver fibrosis (alcoholic and non-alcoholic fatty liver disease, liver cirrhosis, cardiac cirrhosis,cholangitis, portal hypertension, hepatitis-induced fibrosis, viral infection-induced, auto-immune hepatitis, hemochromatosis etc.), pancreatitis, peritonitis, diabetes mellitus type 1 and 2, multiple sclerosis, systemic lupus, systemic sclerosis, arthritis, collagenosis, sarcoidosis, amyloidosis, fibrosis-associated diseases of the genitals and skin diseases (scar formation, etc.).Fibrosis in the context of the cardiovascular disease may be selected from the group consistingof interstitial fibrosis in the context chronic heart failure, replacement fibrosis in the context of myocardial infarction and mixed forms of cardiac fibrosis within other cardiac diseases, such as genetic cardiomyopathies, heart failure with preserved ejection fraction (HFpEF), cardiac valve diseases, Dressler-syndrome, pericarditis, myocarditis, endocarditis, atrial fibrillation, S13211WO / Universität Heidelberg arrhythmias caused by scar tissue such as ventricular tachycardia, cardiac amyloidosis, vasculitis, and atherosclerosis. In one embodiment the LTBP2 expressing fibroblasts are not cancer-associated fibroblasts. Hence, some embodiments refer to a molecule targeting LTBP2 for use in treatment of fibrosis,such as cardiovascular fibrosis, perivascular fibrosis, vascular fibrosis, kidney fibrosis, lungfibrosis and liver fibrosis, atherosclerosis, and stroke. Some embodiments refer to a molecule targeting membrane associated LTBP2 for use in treatment of cardiovascular fibrosis, perivascular fibrosis, vascular fibrosis, atherosclerosis, and stroke. Some specific embodiments refer to a molecule targeting membrane associated LTBP2for use in treatment of cardiovascular fibrosis, perivascular fibrosis, vascular fibrosis.In one embodiment, fibrosis in the context of the cardiovascular disease is selected from the group consisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiac fibrosis.The molecule targeting membrane associated LTBP2 may recognize and bind to the membrane-associated LTBP2 and thereby improve least one echocardiographic parameter, typically after myocardial infarction. The molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) comprises at least one LTBP2 binding domain binding to membrane-associated LTBP2.By the binding of the LTB2 binding domain an interaction between the LTBP2 expressing fibroblasts and an immune cell is established that allows killing of LTBP2 expressing fibroblasts. In one embodiment, the number of activated cardiac fibroblasts is reduced. In one example theLTBP2 binding domain binds an LTBP2 epitope that allows to form an immune synapse betweenthe CAR T cell to mediate killing of LTBP2 expressing fibroblasts. In another example the molecule targeting membrane-associated LTBP2 is a soluble molecule (e.g. an multifunctional antibody, such as a bispecific antibody) that links the LTBP2 expressing fibroblasts and the immune cell to mediate killing of LTBP2 expressing fibroblasts.The LTBP2 binding domain thus binds to an LTBP2 epitope that allows to form an immunesynapse between the LTBP2 expressing fibroblasts and an immune cell to mediate killing of LTBP2 expressing fibroblasts. In some embodiments the molecule targeting membrane bound LTBP2 is engineered to contain at least one of the following: linker sequences, FC-silencing mutations, Fc mutation increasingthe binding to CD16(FcγRIII), modifications to enable inter HC disulfide bridges, modifications toenable LC-HC disulfide bridges, knob-in-hole modifications, and / or purification tags. S13211WO / Universität Heidelberg “Knob-in-hole” modifications are typically modifications in the heavy chain of the antibody enabling specific paring of two different antibody chains, in particular for bispecific antibodies. The “knob“ is a specific residue, typically a bulky amino acid that protrudes from the surface of the antibody structure. Correnspondingly, the “hole” is a complementary modification in the otherheavy chain creating a cavity in the structure that allows the know to fit into this indentation.In one embodiment, the echocardiographic parameter is selected from the group consisting of ejection fraction, fractional area change and end-diastolic volume. In one embodiment, the ejection fraction is left ventricular ejection fraction and / or right ventricular ejection fraction. In one embodiment, the end-diastolic volume is left ventricular end-diastolic volume and / or rightventricular end-diastolic volume. In one embodiment, the ejection fraction may be increased bythe administration of molecule targeting membrane associated LTBP2. In one embodiment, the fractional area change may be increased by the administration of molecule targeting membrane associated LTBP2. In one embodiment, the end-diastolic volume may be decreased by the administration of molecule targeting membrane associated LTBP2.The term “molecule targeting membrane bound LTBP2” refers to a molecule that targets LTBP2which is associated with the extracellular side of the cell membrane, e.g. the extracellular side of a fibroblast. In specific embodiments the molecule is selected from the group consisting of chimeric antigen receptor (CAR), an antibody, a bispecific antibody, a bispecific T cell engager (BiTE), a nanobody and a small molecule. In preferred embodiments, the molecule targetingmembrane bound LTBP2 is selected from the group consisting of CAR, antibody, bispecificantibody, BiTE and nanobody. In a specific embodiment the molecule targeting membrane bound LTBP2 is a CAR. In one embodiment, the molecule targeting membrane bound LTBP2 may comprise a single- chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1 having theamino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ IDNO:2, a CDR3 having the amino acid sequence of SEQ ID NO:3, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO:6. In one embodiment molecule targeting membrane bound LTBP2 may comprise a single-chain variable fragment (scFv) comprising a variable heavy chain having the amino acidsequence that is identical to or 80% identical to SEQ ID NO: 7, and a variable light chain having the amino acid sequence that is identical to or 80% identical to SEQ ID NO:8. In some embodiments the variable heavy chain and the variable light chain might be connected by linker, preferably a peptide linker, more preferably a peptide linker comprising glycine and / or serineresidues. In one embodiment the linker comprises 10 to 20 amino acids, preferably 12 to 18amino acids, more preferably 13 to 17 amino acids such as 15 amino acids. In one embodiment the linker has an amino acid sequence as set out in SEQ ID NO: 9. In a specific embodiment, the molecule targeting membrane bound LTBP2 is a CAR. Table 1. CAR sequences S13211WO / Universität HeidelbergDomain name AA sequence Nucleotide sequenceLTBP2 Vh QGQLQQSGAELARPGASVNLSCKGSGYcagggacagctgcaacagtctggcgccgaacttgctag IFANFGVTWVRQRTGQGLEWIGEIYPGS acctggcgcctctgtgaacctgagctgtaaaggcagcg GNIYYNEKFKGKATLTADKSSSTVYLQFS gctacatcttcgccaacttcggcgtgacctgggtccgaca SLTSEDSAVYFCARSANWGQGTLVTVS gagaacaggacaaggcctggaatggatcggcgagat A ctatcccggctccggcaacatctactacaacgagaagtt caagggcaaagccacactgaccgccgacaagagca (SEQ ID NO: 7) gcagcacagtgtacctgcagttcagcagcctgaccagc gaggatagcgccgtgtacttctgtgccagaagcgccaat tggggccagggcacactggttacagtttctgct (SEQ ID NO: 20)CDR1 GYIFANF (SEQ ID NO: 1) ggctacatcttcgccaacttc(SEQ ID NO: 14)CDR2 YPGSGN (SEQ ID NO: 2) tatcccggctccggcaac(SEQ ID NO: 15)CDR3 SAN (SEQ ID NO: 3) agcgccaat(SEQ ID NO: 16)LTBP2 Vl DVVMTQTPLTLSIAIGQPASISCKSSQSLLGatgtggtcatgacacagacccctctgacactgtctatc HSDGKTYLNWLLQRPGQSPKRLIYLVSK gccatcggacagcctgccagcatcagctgcaagtctag LDPGVPDRFTGSGSGADFTLKISRVEAE ccagagcctgctgcactccgacggcaagacctacctga DLGVYFCWQGTHFPRTFGGGTKLEIK attggctgctgcagaggcccggacagagccccaagag actgatctacctggtgtccaagctggaccctggcgtgccc (SEQ ID NO: 8) gatagattcacaggatctggaagcggagccgacttcac cctgaagattagcagagtggaagccgaggacctgggc gtctacttctgttggcaaggcacacacttccccagaacctt tggcggaggcacaaagctggaaatcaaa (SEQ ID NO: 21)CDR1 KSSQSLLHSDGKTYLN (SEQ ID NO: 4) agtctagccagagcctgctgcactccgacggcaagacctacctgaat (SEQ ID NO: 17)CDR2 LVSKLDP (SEQ ID NO:5) ctggtgtccaagctggaccct(SEQ ID NO: 18)CDR3 WQGTHFPRT (SEQ ID NO: 6) tggcaaggcacacacttccccagaacc(SEQ ID NO: 19)GlySer Linker GGGGSGGGGSGGGGS (SEQ ID NO: 9) ggtggcggaggatctggcggaggtggaagcggcggaggcggatct (SEQ ID NO: 22) S13211WO / Universität Heidelberg hFc (CH2CH3) APELLGGPSVFLFPPKPKDTLMISRTPEVGcacctgaactcctggggggaccgtcagtcttcctcttcc TCVVVDVSHEDPEVKFNWYVDGVEVHN ccccaaaacccaaggacaccctcatgatctcccggac AKTKPREEQYNSTYRVVSVLTVLHQDWL ccctgaggtcacatgcgtggtggtggacgtgagccacg NGKEYKCKVSNKALPAPIEKTISKAKGQP aagaccctgaggtcaagttcaactggtacgtggacggc REPQVYTLPPSRDELTKNQVSLTCLVKG gtggaggtgcataatgccaagacaaagccgcgggag FYPSDIAVEWESNGQPENNYKTTPPVLD gagcagtacaacagcacgtaccgtgtggtcagcgtcct SDGSFFLYSKLTVDKSRWQQGNVFSCS caccgtcctgcaccaggactggctgaatggcaaggagt VMHEALHNHYTQKSLSLSPGK acaagtgcaaggtctccaacaaagccctcccagcccc catcgagaaaaccatctccaaagccaaagggcagcc (SEQ ID NO: 10) ccgagaaccacaggtgtacaccctgcccccaagccgg gatgagctgaccaagaaccaggtcagcctgacctgcct ggtcaaaggcttctatcccagcgacatcgccgtggagtg ggagagcaatgggcagccggagaacaactacaaga ccacgcctcccgtgttggactccgacggctccttcttcctct acagcaagctcaccgtggacaagagcaggtggcagc aggggaacgtcttctcatgctccgtgatgcatgaggctct gcacaaccactacacgcagaagagcctctccctgtctc ccgggaaa (SEQ ID NO: 23) hCD28 FWVLVVVGGVLACYSLLVTVAFIIFWVRSTtttgggtgctggtggtggttggtggagtcctggcttgctat KRSRLLHSDYMNMTPRRPGPTRKHYQP agcttgctagtaacagtggcctttattattttctgggtgagg YAPPRDFAAYRS agtaagaggagcaggctcctgcacagtgactacatga acatgactcccagacgccctggccccacccgcaagca (SEQ ID NO: 11) ttaccagccctatgcccccccacgcgacttcgcagcctat agatct (SEQ ID NO: 24) h4-1BB RFSVVKRGRKKLLYIFKQPFMRPVQTTQCgtttctctgttgttaaacggggcagaaagaagctcctgt EEDGCSCRFPEEEEGGCEL atatattcaaacaaccatttatgagaccagtacaaactac tcaagaggaagatggctgtagctgccgatttccagaag (SEQ ID NO: 12) aagaagaaggaggatgtgaactg (SEQ ID NO: 25) hCD3zeta RVKFSRSADAPAYQQGQNQLYNELNLGAgagtgaagttcagcaggagcgcagacgcccccgcgt RREEYDVLDKRRGRDPEMGGKPRRKN accagcagggccagaaccagctctataacgagctcaa PQEGLYNELQKDKMAEAYSEIGMKGER tctaggacgaagagaggagtacgatgttttggacaaga RRGKGHDGLYQGLSTATKDTYDALHMQ gacgtggccgggaccctgagatggggggaaagccga ALPPR gaaggaagaaccctcaggaaggcctgtacaatgaact gcagaaagataagatggcggaggcctacagtgagatt (SEQ ID NO: 13) gggatgaaaggcgagcgccggaggggcaaggggca cgatggcctttaccagggtctcagtacagccaccaagg acacctacgacgcccttcacatgcaggccctgccccctc gctaa (SEQ ID NO: 26) In one embodiment the molecule targeting membrane bound LTBP2 may comprise a single- chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33 (GYIFANFG), a CDR2 having the amino acid sequenceof SEQ ID NO:34 (IYPGSGNI), a CDR3 having the amino acid sequence of SEQ ID NO:35(ARSAN), a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 36 (QSLLHSDGKTY), a CDR2 having the amino acid sequence LVS and CDR3 having the amino acid sequence of SEQ ID NO:38 (WQGTHFPRT). S13211WO / Universität Heidelberg CARs comprise an extracellular target-binding domain, a transmembrane domain that anchors the CAR to the cell membrane, and one or more endodomains comprising activating domains. The extracellular target-binding domain is typically derived from an antibody and the endodomain comprises signaling modules typically derived from T cell signaling proteins. In apreferred embodiment, the extracellular target-binding domain preferably comprises variableregions from the heavy and light chains of an immunoglobulin configured as a single-chain variable fragment (scFv). The scFv is preferably attached to a hinge region that provides flexibility and transduces signals through an anchoring transmembrane moiety to an intracellular signaling domain. The transmembrane domains originate may for example be from CD8a orCD28.Thus, in some embodiments the CAR may have an extracellular target-binding domain, a transmembrane domain that anchors the CAR to the cell membrane, and one or more activating domains. In some embodiments, the CAR described herein may further include one or more costimulatorydomains.In some embodiments, CAR as described herein may comprise, from N- terminus to C-terminus, the extracellular target-binding domain, the transmembrane domain, and one or more activating domains. In some embodiments, the CAR as described herein may comprise, from N-terminus to C-terminus, the extracellular target-binding domain, the transmembrane domain, one or moreactivating domains, and one or more costimulatory domains.In some embodiments, CAR described herein further comprises a hinge domain that is located between the extracellular target-binding domain and the transmembrane domain. A hinge domain is generally found between two domains of a protein and may allow for flexibility of the protein and movement of one or both of the domains in relation to each other.In some embodiments, the extracellular target-binding domain may be a single-chain antibodyfragment (scFv), which may be derived from a monoclonal antibody described herein. The extracellular target-binding domain may comprise an scFv derived from an anti-LTBP2 antibody described herein. In a specific embodiment, the scFv as described herein includes one, two or three CDRs fromthe heavy chain variable region (e.g., CDR1, CDR2 and / or CDR3); and / or one, two or threeCDRs from the light chain variable region (e.g., CDR1, CDR2 and / or CDR3) provided in Table 1. In some embodiments, the extracellular target-binding domain may further include a linker. The “linker” as used in the context of a scFv refers to a peptide linker that consists of amino acids such as glycine and / or serine residues used alone or in combination, to link variable heavy andvariable light chain regions together. In one embodiment, the flexible polypeptide linker is aGly / Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Ser)n, wherein n is 1, 2, 3, 4, 5, or 6, preferably 3 (SEQ ID NO: 9). S13211WO / Universität Heidelberg In some embodiments, the transmembrane domain may anchor the CAR to the T cell membrane. Transmembrane domains are classified based on the three-dimensional structure of the transmembrane domain. For example, transmembrane domains may form an alpha helix, a complex of more than one alpha helix, a beta-barrel, or any other stable structure capable ofspanning the phospholipid bilayer of a cell.In some embodiments, the activating domain described herein may be involved in triggering cell signaling that leads to a cell activation. Exemplary activating domains include the activating domain of CD3zeta, e.g. as set out in SEQ ID NO: 13. In some embodiments, the costimulatory domains described herein refer to at least a fragmentof a co-stimulatory signaling protein that mediates signal transduction within a cell to induce animmune response such as an effector function. Exemplary costimulatory domains include, but are not limited to, 4-1BB, CD28, 0X40, ICOS, CD40, CD40L, CD27, GITR, HVEM, TIM1, LFA1 or CD2. In some embodiments, the costimulatory domain and the activating domain are for use in CARconstructs disclosed herein that are to be introduced into T cells.In a specific embodiment the CAR may have one or more of the costimulatory domains and one or more activating domains. In a specific embodiment the CAR may have one or more of the costimulatory domains selected from the group consisting of a CD28 domain and a 4-1BB domain and a CD3zeta activatingdomain.In a specific embodiment, the CD28 co-stimulatory domain in the CAR may have a sequence which is identical or at least 80% identical to SEQ ID NO: 11. In a specific embodiment, the 4-1BB co-stimulatory domain in the CAR may have a sequence which is identical or at least 80% identical to SEQ ID NO: 12.In a specific embodiment, the CD3zeta activating domain in the CAR may have a sequencewhich is identical or at least 80% identical to SEQ ID NO: 13. The CAR targeting membrane bound LTBP2 may comprise a single-chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO:2, a CDR3 having theamino acid sequence of SEQ ID NO:3, a variable light chain comprising a CDR1 having theamino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO:6. In one embodiment, the CAR targeting membrane bound LTBP2 may comprise a single-chain variable fragment (scFv) comprising a variable heavy chain having the amino acid sequence that is identical to or 80%identical to SEQ ID NO: 7, and a variable light chain having the amino acid sequence that isidentical to or 80% identical to SEQ ID NO:8. S13211WO / Universität Heidelberg Thus, specific embodiments refer to a CAR comprising from N-terminus to C-terminus: -the scFv as defined herein,- CD28 co-stimulatory domain,- 4-1BB co-stimulatory domain, and- CD3zeta activating domain.Thus, specific embodiments refer to a CAR comprising from N-terminus to C-terminus: -the scFv comprising a variable heavy chain comprising a CDR1 having the aminoacid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO:2, a CDR3 having the amino acid sequence of SAN, a variable light chaincomprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO:6, -CD28 co-stimulatory domain having a sequence which is identical or at least 80%identical to SEQ ID NO: 11,- 4-1BB co-stimulatory domain having a sequence which is identical or at least 80%identical to SEQ ID NO: 12, and -CD3zeta activating domain having have a sequence which is identical or at least 80%identical to SEQ ID NO: 13. In a specific embodiment, the modified cell expressing the molecule targeting membrane bound LTBP2, in particular the CAR, may be a T cell, a NK cell, a dendritic cell, a macrophage, a monocyte, a B cell, a neutrophil, an eosinophil, a basophil, a mast cell, a myeloid derived suppressor cell, a mesenchymal stem cell, a precursor thereof, or a combination.Further aspects relate to a nucleic acid encoding the CAR as described herein: Nucleic acidsequences encoding the relevant domains as described herein are set out in Table 1. In some embodiments, the disclosure features a nucleic acid encoding CAR, wherein the CAR comprises scFv domain. In some specific embodiments, the nucleic acid molecule encoding the scFv described herein may include a nucleotide sequence encoding a variable heavy chain,wherein the variable heavy chain comprises a CDR1 having the nucleotide sequence of SEQ IDNO: 14, a CDR2 having the nucleotide sequence of SEQ ID NO: 15, a CDR3 having the nucleotide sequence of SEQ ID NO: 16, a variable light chain, wherein a variable light chain comprises a CDR1 having the nucleotide sequence of SEQ ID NO: 17, a CDR2 having the nucleotide sequence of SEQ ID NO: 18 and CDR3 having the nucleotide sequence of SEQ IDNO: 19, or a variable heavy chain having a nucleotide sequence identical to or at least 80%identical to the sequence set out in SEQ ID NO: 20, or a variable light chain having a nucleotide sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 21. In some embodiments, the variable heavy chain and the variable light chain described herein might be connected by linker, preferably a peptide linker, more preferably a GlySer Linker. Inone embodiment the linker comprises a nucleotide sequence as set out in SEQ ID NO: 22. S13211WO / Universität Heidelberg In some embodiments, the disclosure features a nucleic acid encoding CAR, wherein the CAR may further comprise one or more of the costimulatory domains and one or more activating domains. In some specific embodiments, the CAR described herein may have one or more of the encodedcostimulatory domains selected from the group consisting of a CD28 domain and a 4-1BBdomain and a CD3zeta activating domain. In some embodiments, the nucleic acid sequence encoding the CD28 co-stimulatory domain may sequence of SEQ ID NO: 24, or a nucleotide sequence with at least 80% identity thereto. In some embodiments, the nucleic acid sequence encoding the 4-1BB co-stimulatory domaincomprises a sequence of SEQ ID NO: 25, or a nucleotide sequence with at least 80% identitythereto. In some embodiments, the nucleic acid sequence encoding the CD3zeta activating domain may comprise nucleotide sequence of SEQ ID NO: 26, or a nucleotide sequence with at least 80% identity thereto. In some embodiments the antibody comprises a single-chain variable fragment (scFv)comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO:2, a CDR3 having the amino acid sequence of SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO:6. In one embodiment molecule targetingmembrane bound LTBP2 may comprise a single-chain variable fragment (scFv) comprising avariable heavy chain having the amino acid sequence that is identical to or 80% identical to SEQ ID NO: 7, and a variable light chain having the amino acid sequence that is identical to or 80% identical to SEQ ID NO:8. In one embodiment the antibody the Fc region of the antibody (CH2CH3) may have an aminoacid sequence that is identical to or 80% identical to SEQ ID NO: 10.Another aspect of the invention refers to a vector comprising the nucleic acid encoding the molecule targeting membrane associated LTBP2, such as a CAR. A “vector” is any molecule or composition that has the ability to carry a nucleic acid sequence into a suitable host cell where synthesis of the encoded polypeptide can take place. Typically, andpreferably, a vector is a nucleic acid that has been engineered, using recombinant DNAtechniques that are known in the art, to incorporate a desired nucleic acid sequence (e.g. a nucleic acid of the invention). The vector may comprise DNA or RNA and / or comprise liposomes. The vector may be a plasmid, shuttle vector, phagemide, cosmid, expression vector, retroviral vector, lentiviral vector, adenoviral vector or particle. A vector may include nucleic acid sequences thatpermit it to replicate in a host cell, such as an origin of replication. A vector may also include oneor more selectable marker genes and other genetic elements known to those of ordinary skill in the art. A vector preferably is an expression vector that includes a nucleic acid according to the present invention operably linked to sequences allowing for the expression of said nucleic acid. S13211WO / Universität Heidelberg Another aspect refers to a molecule targeting membrane associated LTBP2, comprising at least one LTBP2 binding domain binding to membrane associated LTBP2 and at least one engineered immune cell binding domain binding to an immune cell. Typically, such molecule issoluble and links the LTBP2 expressing fibroblast with the immune cell. In some embodimentsthe immune cell is a T cell or a NK cell. Accordingly, some embodiments refer to T cell engagers, in particular bispecific T cell engagers. Thus, some embodiments refer to multispecific antibodies, in particular bispecific antibodieslinking T cells with LTBP2 expressing fibroblast. Exemplary binding domains binding to a T cellare a CD3 binding domain or a CD28 binding domain. Also other surface molecules of a T cell could be used as a target for the cell binding domain binding to an immune cell. Such binding domains may be a scFv, a Fab fragment or a single domain antibody (sdAb), preferably a scFv or a Fab, more preferably a Fab.In one embodiment the immune cell binding domain is an anti-CD28 scFv, an anti-CD28 Fab, ananti CD3ε scFv or anti CD3ε Fab. Accordingly, one embodiment refers to an antibody comprising an anti-CD3ε Fab domain and a anti-LTBP2 Fab domain. Another embodiment refers to an antibody comprising an anti-CD3εscFv domain and an anti-LTBP2 scFv domain.

[0002] S13211WO / Universität Heidelberg Table 2: anti- CD3ε sequences Domain name AA sequenceAnti-CD3ε VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTDKSSSTAYMQLSSLTSE DSAVYYCARYYDDHYCLDYWGQGTTLTVSS (SEQ ID NO: 40) Anti-CD3ε VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLTISGMEAEDAATYYCQQW SSNPFTFGSGTKLEIN (SEQ ID NO: 41) VH-CDR1 GYTFTRYT (SEQ ID NO: 42)VH-CDR2 INPSRGYT (SEQ ID NO: 43)VH-CDR3 ARYYDDHYCLDY (SEQ ID NO: 44)VL-CDR1 SSVSY(SEQ ID NO: 45)VL-CDR2 DTSVL-CDR3 QQWSSNPFT (SEQ ID NO: 47)Table 2 provides exemplary sequences comprised by an anti-CD3ε binding molecule.Accordingly, one embodiment refers to a bispecific antigen binding molecule comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 42, a CDR2 having the amino acid sequence of SEQ ID NO: 43, a CDR3 having the amino acidsequence of SEQ ID NO: 44, a variable light chain comprising a CDR1 having the amino acidsequence of SEQ ID NO: 45, a CDR2 having the amino acid sequence DTS and CDR3 having the amino acid sequence of SEQ ID NO: 47, and a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acidsequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence ofSEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6. Another embodiment refers to a bispecific antigen binding molecule comprising a variable heavychain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 42, a CDR2 havingthe amino acid sequence of SEQ ID NO: 43, a CDR3 having the amino acid sequence of SEQ ID NO: 44, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 45, a CDR2 having the amino acid sequence DTS and CDR3 having the amino acid sequence of SEQ ID NO: 47, and S13211WO / Universität Heidelberg a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ ID NO: 35, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 36, a CDR2 having the amino acid sequence LVS and CDR3 havingthe amino acid sequence of SEQ ID NO: 38.Another embodiment refers to a bispecific antigen binding molecule comprising a variable heavychain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 42, a CDR2 havingthe amino acid sequence of SEQ ID NO: 43, a CDR3 having the amino acid sequence of SEQ ID NO: 44, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 45, a CDR2 having the amino acid sequence DTS and CDR3 having the amino acid sequence of SEQ ID NO: 47 andan a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO:1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6. Another embodiment refers to a bispecific antigen binding molecule comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 42, a CDR2 having the amino acid sequence of SEQ ID NO: 43, a CDR3 having the amino acid sequence of SEQ ID NO: 44, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ IDNO: 45, a CDR2 having the amino acid sequence DTS and CDR3 having the amino acidsequence of SEQ ID NO: 47 and an comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ ID NO: 35, a variable light chain comprising a CDR1 havingthe amino acid sequence of SEQ ID NO: 36, a CDR2 having the amino acid sequence LVS andCDR3 having the amino acid sequence of SEQ ID NO: 38. In one embodiment the bispecific antigen binding molecule may comprise a variable heavy chain having the amino acid sequence that is at least 80% identical to SEQ ID NO: 7, and a variablelight chain having the amino acid sequence that is at least 80% identical to SEQ ID NO:8targeting membrane bound LTBP2 and a variable heavy chain having the amino acid sequence that is at least 80% identical to SEQ ID NO: 40, and a variable light chain having the amino acid sequence that is at least 80% identical to SEQ ID NO:41 targeting CD3ε. In one embodiment the bispecific antigen binding molecule may comprise a variable heavy chainhaving the amino acid sequence that is at least 90% identical to SEQ ID NO: 7, and a variablelight chain having the amino acid sequence that is at least 90% identical to SEQ ID NO:8 targeting membrane bound LTBP2 and a variable heavy chain having the amino acid sequence S13211WO / Universität Heidelberg that is at least 90% identical to SEQ ID NO: 40, and a variable light chain having the amino acid sequence that is at least 90% identical to SEQ ID NO:41 targeting CD3ε. In one embodiment the bispecific antigen binding molecule may comprise a variable heavy chain having the amino acid sequence of SEQ ID NO: 7, and a variable light chain having the aminoacid sequence of SEQ ID NO:8 targeting membrane bound LTBP2 and a variable heavy chainhaving the amino acid sequence of SEQ ID NO: 40, and a variable light chain having the amino acid sequence of SEQ ID NO:41 targeting CD3ε. Accordingly, some embodiments refer to NK cell engagers. Thus some embodiments referantibodies linking NK cells with LTBP2 expressing fibroblast.Exemplary binding domains binding to a NK cell are engineered Fc domain that interacts with the CD16 domain of NK cells. For example the engineered Fc domain may contain mutations that enhance interaction of the Fc domain with the NK cell (see for example the S239D and I332E mutations as described in Gauthier et al., 2019, Cell 177, 1701–1713). An example of an engineered Fc domain is set out in the following sequence (SEQ ID NO: 39): EPKSSDKTHTCPPCPAPELLGGPDVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPLPEEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Alternative embodiments refer to antibodies linking NK cells with LTBP2 expressing fibroblasts, relate to multispecific antibodies, in particular bispecific antibodies comprising a CD16 bindingdomain such as a scFv, a Fab fragment or a single domain antibody (sdAb), preferably a scFv ora Fab, more preferably a Fab. The molecule targeting membrane associated LTBP2 may be an multifunctional protein, such as a bifunctional or a trifunctional protein. For example, the molecule targeting membrane associatedLTBP2 may be a multispecific antibody, such as a bispecific antibody.In one embodiment, the bifunctional protein is an anti-LTBP2 / anti-hCD3ɛ bispecific bivalent antibody comprising three domains forming a heterotrimer, preferably comprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100% identical to the amino acidsequence of SEQ ID NO: 53, an amino acid sequence that is at least 80%, at least 90%, at least95% or 100% identical to the amino acid sequence of SEQ ID NO: 54, and an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100% identical to the amino acid sequence of SEQ ID NO: 55. In one embodiment, the bifunctional protein is an anti-LTBP2 / anti-hCD3ɛ bispecific antibodycomprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100%identical to the amino acid sequence of SEQ ID NO: 59. S13211WO / Universität Heidelberg In one embodiment, the bifunctional protein is an anti-LTBP2 / anti-mCD3ɛ bispecific antibody comprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100% identical to the amino acid sequence of SEQ ID NO: 63. In one embodiment, the bifunctional protein is an anti-LTBP2 / anti-mCD28 bispecific antibodycomprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100%identical to the amino acid sequence of SEQ ID NO: 67. In one embodiment, the bifunctional protein is an anti-LTBP2 bifunctional NK-cell engaging antibody (anti human) comprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100% identical to the amino acid sequence of SEQ ID NO: 61.In one embodiment, the bifunctional protein is an anti-LTBP2 bifunctional NK-cell engagingantibody (anti mouse) comprising an amino acid sequence that is at least 80%, at least 90%, at least 95% or 100% identical to the amino acid sequence of SEQ ID NO: 65. In one embodiment, the bifunctional protein is an anti-LTBP2 bifunctional NK-cell engaging antibody (anti mouse) comprising an comprising an Fc domain according to SEQ ID NO: 39 anda variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, aCDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6. In one embodiment, the bifunctional protein is an anti-LTBP2 bifunctional NK-cell engaging antibody (anti mouse) comprising an comprising an Fc domain according to SEQ ID NO: 39 and a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acidsequence of SEQ ID NO: 35, a variable light chain comprising a CDR1 having the amino acidsequence of SEQ ID NO: 36, a CDR2 having the amino acid sequence LVS and CDR3 having the amino acid sequence of SEQ ID NO: 38. Another aspect refers to a pharmaceutical composition comprising the molecule targetingmembrane associated LTBP2 as described herein together with a pharmaceutically acceptablecarrier. The term “composition” as used herein refers to a mixture comprising a therapeutically effective amount of the agent according to the present invention and one or more excipients. The term“excipient” as used herein may also be referred to as “pharmaceutically acceptable carrier”, or“pharmaceutically acceptable excipient,” “pharmaceutically acceptable diluent,”, or “pharmaceutically acceptable vehicle,” used interchangeably herein, refer to a non-toxic solid, semisolid or liquid filler, diluent, encapsulating material or formulation auxiliary of any conventional type. S13211WO / Universität Heidelberg A pharmaceutically acceptable carrier is essentially non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. A pharmaceutically acceptable carrier will not inhibit otherwise adversely affect the function of the agent according to the present invention. Suitable carriers include, but are not limited to water,dextrose, glycerol, saline, ethanol, and any combination thereof. The carrier can containadditional agents such as wetting or emulsifying agents, pH buffering agents, or adjuvants, which enhance the effectiveness of the formulation. As used herein, the term “excipient” refers to an inert substance which is commonly used as adiluent, vehicle, preservative, binder, or stabilizing agent, and includes, but is not limited to,proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate, etc.), surfactants (e.g., SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). Another aspect refers to a cell modified to express the molecule targeting membrane associated LTBP2, such as a CAR. In some embodiments, the cell can be a T cell, a NK cell, a dendritic cell, a macrophage, a monocyte, a B cell, a neutrophil, an eosinophil, a basophil, a mast cell, a myeloid derived suppressor cell, a mesenchymal stem cell, a precursor thereof, or acombination.In one embodiment the cell is a lymphocyte. The lymphocyte may be for example a T cell or NK cell. In one embodiment the cell is myeloid cell. The myeloid cell may be for example a macrophage or a monocyte.Another aspect of the invention refers to the use of molecule targeting membrane associatedlatent-transforming growth factor beta-binding protein 2 (LTBP2) in diagnosing a disease associated with LTBP2 expressing fibroblasts. Further Embodiments The invention is further described by the following embodiments: Embodiment 1: Molecule targeting membrane associated latent-transforming growth factor beta- binding protein 2 (LTBP2) for use in treatment or prevention of a disease associated with LTBP2expressing fibroblasts.Embodiment 2: Molecule targeting membrane associated LTBP2 according to embodiment 1, wherein LTBP2 is associated to the membrane in the LTBP2 expressing fibroblasts. S13211WO / Universität Heidelberg Embodiment 3: Molecule targeting membrane associated LTBP2 for use according to embodiment 1 or 2, wherein the molecule targeting membrane associated LTBP2 is selected from the group consisting of a chimeric antigen receptor (CAR), an antibody, a bispecific antibody, a bispecific T cell engager (BiTE), a nanobody and a small molecule. Embodiment 4: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding embodiments, wherein the molecule targeting membrane associated LTBP2 comprises a single-chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acidsequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chaincomprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6.Embodiment 5: Molecule targeting membrane associated LTBP2 for use according to any one ofthe preceding embodiments, wherein the molecule targeting membrane associated LTBP2 comprises a single-chain variable fragment (scFv) comprising a variable heavy chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 7.Embodiment 6: Molecule targeting membrane associated LTBP2 for use according to any one ofthe preceding embodiments, wherein the molecule targeting membrane associated LTBP2 comprises a single-chain variable fragment (scFv) comprising a variable light chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 8.Embodiment 7: Molecule targeting membrane associated LTBP2 for use according to any one ofthe preceding embodiments, wherein the molecule targeting membrane associated LTBP2 is a CAR comprising from N-terminus to C-terminus: -the scFv as defined in embodiments 4 to 6,- optionally at least one co-stimulatory domain, and- an activating domainEmbodiment 8: Molecule targeting membrane associated LTBP2 for use according to embodiment 7, wherein the at least one co-stimulatory domain is selected from the group consisting of CD28 and 4-1BB. Embodiment 9: Molecule targeting membrane associated LTBP2 for use according to embodiment 7 or 8, wherein the activating domain is CD3zeta. Embodiment 10: Molecule targeting membrane associated LTBP2 for use according to any oneof the preceding embodiments, wherein the molecule targeting membrane bound LTBP2 is aCAR comprising from N-terminus to C-terminus: -the scFv as defined in embodiments 4 to 6,- a CD28 co-stimulatory domain,- a 4-1BB co-stimulatory domain, and S13211WO / Universität Heidelberg -a CD3zeta activating domain.Embodiment 11: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding embodiments, wherein the CD28 co-stimulatory domain has a sequenceidentical to or at least 80% identical to SEQ ID NO.: 11.Embodiment 12: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding embodiments, wherein the 4-1BB co-stimulatory domain has a sequence identical to or at least 80% identical to SEQ ID NO.: 12. Embodiment 13: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding embodiments, wherein the CD3zeta activating domain has a sequence which is identical to or at least 80% identical to SEQ ID NO.: 13.Embodiment 14: Molecule targeting membrane associated LTBP2 for use according to any oneof the preceding embodiments, wherein the disease associated with LTBP2 expressing fibroblasts is fibrosis. Embodiment 15: Molecule targeting membrane associated LTBP2 for use according to any oneof the preceding embodiments, wherein the disease associated with LTBP2 expressingfibroblasts is selected from the group consisting of fibrosis in the context of the cardiovascular disease, kidney fibrosis, lung fibrosis and liver fibrosis. Embodiment 16: Molecule targeting membrane associated LTBP2 for use according toembodiment 15, wherein fibrosis in the context of the cardiovascular disease is selected fromthe group consisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiac fibrosis. Embodiment 17: Molecule targeting membrane associated LTBP2 for use according to any oneof the preceding embodiments, wherein at least one echocardiographic parameter is improved.Embodiment 18: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding embodiments, wherein at least one echocardiographic parameter is improved after myocardial infarction. Embodiment 19: Molecule targeting membrane associated LTBP2 for use according to any one of embodiment 17 or 18, wherein at least one echocardiographic parameter is selected from the group consisting of ejection fraction, fractional area change and end-diastolic volume.Embodiment 20: Molecule targeting membrane associated LTBP2 for use according toembodiment 19, wherein the ejection fraction is left ventricular ejection fraction and / or right ventricular ejection fraction. S13211WO / Universität Heidelberg Embodiment 21: Molecule targeting membrane associated LTBP2 for use according to embodiment 19 or 20, wherein the end-diastolic volume is left ventricular end-diastolic volume and / or right ventricular end-diastolic volume.Embodiment 22: CAR targeting LTBP2, comprising from N-terminus to C-terminus:- the scFv as defined in embodiments 4 to 6,- at least one co-stimulatory domain, and- an activating domain.Embodiment 23: CAR according to embodiment 16, wherein the at least one co-stimulatorydomain is selected from the group consisting of CD28 and 4-1BB. Embodiment 24: CAR according to embodiment 16 or 17, wherein the activating domain is CD3zeta. Embodiment 25: CAR according to any one of embodiments 226 to 24, wherein the molecule targeting membrane associated LTBP2 is a CAR comprising from N-terminus to C-terminus: -the scFv as defined in embodiments 4 to 6,- CD28 co-stimulatory domain,- 4-1BB co-stimulatory domain, and- CD3zeta activating domain.Embodiment 26: CAR to any one of embodiments 22 to 25, wherein the CD28 co-stimulatory domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO.:11.Embodiment 27: CAR according to any one of the embodiments 22 to 26, wherein the 4-1BB co- stimulatory domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO.: 12. Embodiment 28: CAR according to any one of the embodiments 22 to 27, wherein the CD3zeta activating domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO.: 13.Embodiment 29: Nucleic acid encoding the CAR according to any one of embodiments 22 to 28.Embodiment 30: Vector comprising the nucleic acid according to claim 29. Embodiment 31: Cell modified to express the CAR according to any one of embodiments 22 to28.Embodiment 32: Cell according to embodiment 31, wherein the cell is a leukocyte, or a myeloid cell. S13211WO / Universität Heidelberg Embodiment 33: Cell according to embodiment 32, wherein the leukocyte is a lymphocyte, wherein the lymphocytes is preferably a T cell or NK cell. Embodiment 34: Cell according to embodiment 31, wherein the myeloid cell is a macrophage ora monocyte.Embodiment 35: Use of molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) in diagnosing a disease associated with LTBP2 expressing fibroblasts, fibrosis, chronic heart failure, and / or myocardial infarction. Embodiment 36: Molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) for use in treatment or prevention of fibrosis, chronic heart failure, and / or myocardial infarction.The application comprises further the following items:1. Molecule targeting membrane associated latent-transforming growth factor beta-bindingprotein 2 (LTBP2) for use in treatment of a disease associated with LTBP2 expressing fibroblasts. 2. Molecule targeting membrane associated LTBP2 according to item 1, wherein LTBP2 isassociated to the membrane in the LTBP2 expressing fibroblasts. 3. Molecule targeting membrane associated LTBP2 for use according to item 1 or 2,wherein the molecule targeting membrane associated LTBP2 is selected from the groupconsisting of a chimeric antigen receptor (CAR), an antibody, a bispecific antibody, a bispecific T cell engager (BiTE), a nanobody and a small molecule. 4. Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding items, wherein the molecule targeting membrane associated LTBP2comprises a single-chain variable fragment (scFv) comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO:2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 havingthe amino acid sequence of SEQ ID NO:6. 5. Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding items, wherein the molecule targeting membrane associated LTBP2 comprises a single-chain variable fragment (scFv) comprising a variable heavy chainhaving a sequence identical to the sequence or at least 80% identical to the sequence set out in SEQ ID NO: 7; and a variable light chain having a sequence identical to the sequence or at least 80% identical to the sequence set out in SEQ ID NO: 8. S13211WO / Universität Heidelberg6. Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding items, wherein the molecule targeting membrane associated LTBP2 is a CAR comprising from N-terminus to C-terminus: ^the scFv as defined in items 4 to 5,^ optionally at least one co-stimulatory domain, and^ an activating domain.7. Molecule targeting membrane associated LTBP2 for use according to item 6, wherein theat least one co-stimulatory domain is selected from the group consisting of CD28 and 4-1BB; and wherein optionally the activating domain is CD3zeta.8. Molecule targeting membrane associated LTBP2 for use according to item 7, wherein theCD28 co-stimulatory domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO: 11; the 4-1BB co-stimulatory domain has a sequenceidentical to or at least 80% identical to SEQ ID NO: 12 and the CD3zeta activating domain has a sequence identical to or at least 80% identical to SEQ ID NO: 13.9. Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding items, wherein the disease associated with LTBP2 expressing fibroblasts isfibrosis, optionally wherein the disease associated with LTBP2 expressing fibroblasts is selected from the group consisting of fibrosis in the context of the cardiovascular disease, kidney fibrosis, lung fibrosis and liver fibrosis, optionally wherein fibrosis in the context of the cardiovascular disease is selected from the group consisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiacfibrosis.10. CAR targeting LTBP2, comprising from N-terminus to C-terminus:^ the scFv as defined in items 4 to 4,^ at least one co-stimulatory domain, and^ an activating domain.11. CAR according to item 10, wherein the at least one co-stimulatory domain is selectedfrom the group consisting of CD28 and 4-1BB; and the activating domain is CD3zeta.12. CAR to any one of items 10 to 11, wherein the CD28 co-stimulatory domain has asequence identical or at least 80% identical to SEQ ID NO.: 11; the 4-1BB co-stimulatory domain has a sequence identical or at least 80% identical to SEQ ID NO.: 12 and the CD3zeta activating domain has a sequence identical or least 80% identical to SEQ ID NO.: 13.13. Nucleic acid encoding the CAR according to any one of items 10 to 12. S13211WO / Universität Heidelberg 14. Cell modified to express the CAR according to any one of items 10 to 12, whereinoptionally the cell is a leukocyte, or a myeloid cell. 15. Use of molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) in diagnosing a disease associated with LTBP2 expressingfibroblasts. ExamplesThe inventors have identified the matrix protein latent-transforming growth factor beta-bindingprotein 2 (LTBP2) as a promising therapeutic target using an unbiased bioinformatic meta- analysis. This identification is based on the strong upregulation of LTBP2 at both the RNA and protein levels following myocardial infarction induction in mice, caused by both permanent and transient LAD (left anterior descending artery) occlusion (Fig.1). The expression is restricted tothe fibrotic zone / scar and is not significantly expressed in healthy tissue (Fig. 1). LTBP2expression correlates substantially with disease-driving fibroblast activation, however it is not primarily expressed in the early and vulnerable time points after permanent LAD occlusion myocardial infarction, but at later time points. This makes it a promising therapeutic target, as ruptures as an undesirable consequence of therapy can be avoided. The inventors have demonstrated that further re-analyses of CITE-Seq protein expression datasets from several human organ donors show a very good specificity of LTBP2 for fibroblasts in the human heart (Fig.2A). In addition, the inventors were able to identify LTBP2-expressing fibroblasts as activated fibroblasts (Fig.2B). These fibroblasts were hardly found in the tissue ofheart-healthy organ donors, but are strongly upregulated in acute myocardial infarction, ischemiccardiomyopathy, and somewhat less strongly in non-ischemic cardiomyopathy in patients (Fig. 2C). Thus, the research of the scientists represents LTBP2 as an ideal therapeutic target and marker for pathologic fibroblast activation.It was exhibited by in vitro studies that the mechanistic investigation of LTBP2 expression usinghuman ventricular fibroblasts analyzed shows a TGFβ-induced upregulation of LTBP2 expression (Fig.3). As a central regulator, TGFβ is significantly involved in the development and maintenance of pathological fibrosis processes in many organs. The scientists were able to show that LTBP2 is associated with the plasma membrane of human fibroblasts after stimulationwith TGFβ (Fig. 3B). The immunofluorescence results were confirmed in flow cytometryexperiments of living, non-permeabilized fibroblasts (Fig.3D). Therefore, LTBP2 is considered to be readily accessible for extracellular therapeutic strategies. The biological and pathomechanistic properties of LTBP2 as described in Fig.3 suggest a greattherapeutic potential as a fibrosis-regulating therapeutic target. Ablation or modulation ofactivated fibroblasts expressing LTBP2 could effectively reduce or prevent pathologic fibrosis. In order to utilize this, the scientists validated a specific antibody sequence and used it to design and generate "chimeric antigen receptor T cells" (CAR-T cells) (Fig.4). Using an adenovirus S13211WO / Universität Heidelberg vector, the anti-LTBP2 CAR sequence was introduced into T cells, which were propagated and examined with regard to their function. No unwanted, excessive T cell activation (tonic signaling) and good expression of the CAR receptor on the T cell surface were observed. Functional analyses with human cardiac fibroblasts showed very good specificity and successful applicationfor killing activated fibroblasts in vitro (Fig. 4B). The anti-LTBP2 CAR-T cells were also used in invivo experiments to assess the systemic effects. The first preliminary results of the anti-LTBP2- CAR-T administration at day 6 following experimental myocardial infarction in mice showed an improvement of the cardiac function by amelioration of several echocardiographic parameters, such as left ventricular ejection fraction, fractional area change and left ventricular end-diastolicvolume (Fig.4C) indicating a beneficial effect such as preventing the progression of heart failure.The first in vivo application revealed a safety of the treatment including a long-term follow-up until day 56 after myocardial infarction. As LTBP2 was reported to be expressed in the aortic tissue, it was investigated whether the anti-LTBP2 CAR T cell therapy might induce relevant side effects such as aortic dilation or aneurysm. However, no dilation or aneurysm of the aortic sinusor ascending aorta were observed and no animal died after CAR-T cell injection indicating a verygood tolerance and safety of the treatment. In the following studies, the therapeutic and functional effects of anti-LTBP2 CAR therapy will be further investigated using histology and molecular biology methods. Further optimization of the immunotherapy will follow, including the use of monoclonal antibodies, bispecific antibodies, bispecific T cell engagers (BiTES), LTBP2-bound TGFβ traps, in vivo generated (e.g. mRNA-based) CARs or other upcoming technologiesthat relate on extracellular binding and targeting of LTBP2-expressing stromal cells. MethodsIn vivo experimentsAnimals All animal experiments were conducted in agreement with the animal welfare guidelines and German national laws. All animal procedures and study protocols were authorized and approvedby the responsible authority (permit No. G-297 / 21, Regierungspräsidium Karlsruhe, Baden-Württemberg, Germany). C57BL / 6N female mice, obtained from Janvier Labs, were used at an age of 10-12 weeks. Minimal-invasive myocardial infarction:Acute myocardial infarction (MI) was induced by minimal-invasive permanent or transient(ischemia-reperfusion injury) occlusion of the left-descending coronary artery (LAD), as described and published previously [2]. Ischemia reperfusion myocardial infarctions were induced in a similar way, without applying electrocoagulation and ischemia induced for 45min. The infarct size was determined by troponin T measurements from blood at 24h post infarct.Sham control mice underwent the same procedure except for the LAD occlusion.Echocardiographic measurements: For the monitoring of cardiac function after MI, transthoracic echocardiography was performed on a VisualSonics Vevo 2100 system equipped with MS550 transducer (Visual Sonics). Awake S13211WO / Universität Heidelberg and narcotized echocardiography was performed by induction (4 vol%) and short maintenance (0.5–1.5 vol%) of isoflurane anesthesia. Left ventricular (LV) parasternal long axis and short axis views at the mid-papillary muscle level were acquired. LV end-diastolic volume (LVEDV), fractional area change (FAC), LV fractional shortening (FS) and LV ejection fraction (LVEF) wereobtained next to aortic sinus and ascending aorta diameters.Therapeutic application: Treatment groups were randomized and matched according to infarct size as determined byblood troponin T levels and LVEF at day 1. At day 6 post MI, a total number of 500,000 anti-LTBP2 murine CAR-T cells or PBS as control were injected in a volume of 100uL via the tail vein. The CAR-T cell treatment was very well tolerated by all treated animals and none of them died prematurely following injection. Echocardiography was performed at baseline, day 1, 3, 7, 14, 28 and 31 after MI to access cardiac function. RNA expression analysis: After sacrificing the animals by cervical dislocation, the still beating heart was directly placed in ice-cold HBSS where atria and large vessels were removed and the remote and infarct zone of the heart dissected and immediately placed into liquid nitrogen and stored at -80°. The tissuewas homogenized and total RNA isolated using a RNeasy kit (Qiagen). Quantification andquality controls were performed by DS-11 spectrophotometer (DeNovix). RNA was reverse transcribed using the Revert Aid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific) and SYBR Green (Bio-Rad Laboratories) was used for quantitative real-time PCR (RT-qPCR) according to the manufacturer’s instructions. A ViiA 7 Real-Time PCR System for 384-well plateswas used and analysis performed by normalizing relative changes to hypoxanthinephosphoribosyltransferase (Hprt) mRNA using the 2ΔΔCt method. Fold change mRNA levels are expressed relative to the mean of the indicated control conditions (set to 1). The following primer sequences were used: Target gene (mouse) Forward Primer Reverse PrimerLtbp2 AACAGCACCAACCACTGTATCCCTGGCATTCTGAGGGTCAAA SEQ ID NO: 29 SEQ ID NO: 30 Hprt GTCAACGGGGGACATAAAAGTGCATTGTTTTACCAGTGTCAA SEQ ID NO: 31 SEQ ID NO: 32 Immunofluorescence staining of tissue sections: For immunofluorescence staining, in TissueTek embedded and frozen heart-derived cryostat sections were fixed in 3,7% PFA, permeabilized for 20 min (0,5 % Tween 20 in PBS), blockedfor 1h (5% BSA, 0,1 % Tween 20 in PBS), and stained overnight with anti-LTBP2 (1:200,Biorybt: orb628535) and the following day for one hour with donkey anti-rabbit AlexaFluor647 (1:400, Abcam: Ab150075) secondary antibody. Nuclei were stained with a DAPI solution (300 nM) and cell membranes such as unspecific collagen staining by a wheat germ agglutinin (WGA) solution (5 µg / ml). The stained sections were mounted, covered and images captured of the S13211WO / Universität Heidelberg whole sections in an automated way using a slide-scanner (Axioscan 7, Zeiss) fluorescence microscope. Quantification of the fluorescence area and intensity was performed by QuPath 0.4.4 software. In vitro experiments Human fibroblast stimulation: In vitro experiments included human primary cardiac ventricular fibroblasts (Lonza, #CC-2904 orCell Applications, 306V-05a) at passage 5-8 cultured according to the manufacturer`srecommendations using respective basal and growth media. The in vitro experimental design is depicted in Figure 3A. Cells were plated at 30,000 cells / well in 12-well plates or 3,500 cells / well in 96-well plates and starved in basal medium for 24h. Fibroblasts were stimulated by growth medium and 1-40 ng / ml TGFβ1 (Sigma-Aldrich, T-7039) or respective volumes of PBS ascontrol.48h after stimulation the cells were harvested for RNA isolation, immunofluorescencestaining or flow cytometry. Immunofluorescence staining of human fibroblasts in vitro: Human cardiac fibroblasts were cultured and treated as described above. Cells were plated onautoclaved cover slips in 12-well plates. After the stimulation, the cover slips were washed andstained using anti-LTBP2 antibody (1:200, Biorybt: orb628535) and respective donkey anti- mouse AlexaFluor647 (1:400) secondary antibody, PKH67 (2x10-6 M, Sigma Aldrich) for membrane staining and DAPI. Importantly, no permeabilization reagents were used to restrict the fluorescence signal to membrane-associated LTBP2. Imaging and image processing wereperformed on a Leica SP8 confocal microscope allowing to generate 3D images of cardiacfibroblasts and LTBP2 localization. Flow cytometry: Cultured human fibroblasts were detached by using accutase solution (Sigma Aldrich). Cellswere washed, counted and diluted to 100µl per 10x106 cells prior staining. An additional controlsample was used for unstained and fluorescence-minus-one gating controls. Cells were kept on ice, washed and resuspended for 1:100 antibody staining. For all samples, Fc receptor blocking was performed for 10 min prior to fluorescent antibody staining using an anti-LTBP2 (5D7) antibody and a secondary anti-mouse AlexaFluor594 antibody (abcam, ab150108)subsequently. Gating strategies and representative plots are presented in figure 3D. Flowcytometry was performed on a FACSCelesta (BD) and data analysis conducted by FlowJo (BD) software. CAR-T generation:LTBP2 specific CAR-T cells were generated by lentiviral transduction of the pRRL-cPPT-LTBP2scFv-hFc-hCD28-h4-1BB-hCD3zeta-WPRE expression cassette. A second-generation packaging system was used to generate lentiviral particles. Packaging plasmids pMD2.G and pCMVR8.74 were obtained from Addgene (plasmid # 12259 and # 22036). HEK293T cells were seeded onto 15cm plates 2 days before transfection (6x106 cells / plate, 5 plates comprise one S13211WO / Universität Heidelberg stack). Transfection of cells was done using 7.5mM PEI in 150mM NaCl and with equimolar amounts of plasmids (per stack: 112.5µg pRRLSIN – 39.5µg pMD2.G – 73µg pCMVR8.74). One day after transfection, the medium was replaced with 14ml of RPMI1640 (w / o phenol red, w / o FCS) per plate. Two days after transfection, culture supernatant (70ml per stack) was harvested,filtered (45µm PVDF filter membranes), and concentrated using sterilized Centricon® Plus-70Ultracel® PL-100 devices (Merck). Lentiviral particles from one production were pooled and stored at -80°C. Determination of infectious titer was calculated as described previously [3] by FACS measurement of CAR expression.Primary human CD3 positive T cells were derived from healthy donor PBMCs by using thehuman Pan T cell Isolation Kit (Miltenyi Biotec) according to the manufacturer´s instructions. T cells were activated for 48h by TransAct™ in TexMACS™ medium at a density of 1 Mio cells / ml containing IL-7 (0.1 mg / ml) and IL-15 (0.1 mg / ml) (all from Miltenyi Biotec). Transduction of primary CD3+ cells was done by incubation with concentrated lentiviral supernatant at day 2after activation with a MOI of 5 in TexMACS™ Medium containing IL-7 and IL-15. Determinationof CAR expression was done 48h after transduction by FACS using the APC conjugated anti- CD3 antibody UCHT1 (BD Biosciences) and a PE conjugated polyclonal anti-Fc F(ab’)2 fragment (Jackson ImmunoResearch), respectively.Murine T cells were obtained from freshly isolated spleens squeezed through a cell strainer.Erythrocytes were lysed with ammonium chloride solution and CD3+ cells were isolated with the Pan T cell Isolation Kit II (Miltenyi Biotec). Cells were cultivated in 12 well plates (5x10E6 cells / well) with RPMI-1640 medium supplemented with 10% FCS, 1% PS as well as anti-CD3 antibody (2μg / mL), anti-CD28 antibody (1μg / mL), IL-2 (100IU / mL), L-glutamine (2mM), ß-mercaptoethanol (50μM) and non-essential amino acids (1x). For the generation of murine CAR-Ts, electroporation was performed two days after activation using the Neon™-Transfection device (ThermoFisher) at 1900V, 3 pulses, 20ms, followed by one day of cultivation in mouse T cell media before flow cytometry, as reported previously [4].Therapeutic use of bispecific antibody constructs targeting LTBP2We designed several bispecific antibody constructs (bsMAbs) targeting extracellular murine or human “Latent-transforming growth factor beta-binding protein 2” (LTBP2). Herein we report novel results showing the successful application of human and murine antibody constructs. Furthermore, we validated the functional relevance of LTBP2 in multiple murine disease models,including liver fibrosis and myocardial ischemia-reperfusion injury.Human anti-LTBP2 bispecific constructs: Three different human constructs were generated characterized by the following proteinsequences and structural designs. All of them bind to human LTBP2. The first construct representsa neutralizing CD3 co-transfected anti-LTBP2 antibody (No.: 15199.1, Fig.5A) and the following two have a quadruple antibody structure and do also bind to CD3 (No.: 15452.1, Fig.5B) on T- cells or CD16 (No.: 15517.1, Fig. 5C) on natural-killer cells and other Fc-receptor expressing S13211WO / Universität Heidelberg immune cells, respectively. The same LTBP2 target-specific sequence was used as for the the CAR-T cell generation described above. We validated the above noted human anti-LTBP2 bispecific constructs in vitro and in an “ex-vivo”model using living human myocardial slices (see e.g. Watson, S.A. et al. Nat Commun 10, 2168(2019).). Unless stated differently, the same methods were used as described above (such as immunofluorescence including all antibodies used). First, we performed killing-assays using the xCELLigence (Agilent) platform, which enables a real-time measurement of cell viability by calculating a cell-index from impendence changes on theplate bottom (Fig. 6A). We seeded 15.000 commercially available primary human ventricular cardiac fibroblasts per well (passage 5-8, Cell applications) into a gold microelectrode 96-well plate (E-plate, Agilent) and allowed them to settle down and stabilize for 8 hours within the platform placed into a cell culture incubator at 38°C and 5% CO2. The day before, T or NK cells wereisolated from healthy donors using a MACS microbead kits for untouched human T or NK cellisolation (Miltenyi Biotec) and T cells were stimulated for 24h with interleukin-2, NK cells with interleukin-15 for 24h and kept in culture using respective T or NK cell media (Fig.6A). Either T-cells (100.000 / well) or NK cells (25.000 / well) were added to the fibroblasts next to the matching T or NK cell engaging antibody constructs or PBS as control (Fibroblasts + NK cells;Fibroblasts + T-cells). Changes in cell viability were recorded in real-time to measure the killingefficiency and analysed up to 80h. Treatment with the neutralizing anti-LTBP2 antibody (No.: 15199.1) incubated with NK cells revealed a reduced proliferation and suggests a killing at all used concentrations (Fig. 6B). Application of the bispecific antibody binding CD3 (No.: 15452.1) showed a dose-dependent killingof fibroblasts after incubation with heterologous T cells (Fig. 6C). Finally, the bispecific antibodyengaging with CD16 (No.: 15517.1) and incubation with NK cells was also able to reduce the proliferation of fibroblasts in a dose-dependent manner (Fig.6D). We observed that activated NK cells induced a proliferation whereas heterologous T cells a reduction of fibroblasts within the xCELLigence assay wells, however all constructs showed effective reduction of fibroblasts, giventhat not all of them do express LTBP2.Patients undergoing left-ventricular assist-device (LVAD) implantation show high amounts of LTBP2-accumulation within the diseased, fibrotic heart areas. Such patients were included into the study after giving written informed consent (Ethic protocol Nr.: S-143-2024, Medical Faculty ofthe University of Heidelberg). Blood was drawn by venipuncture into EDTA-monovettes one ortwo days prior to planned surgery for T and NK cell isolation. Cells were isolated as described above and kept in cell culture until surgery and generation of the myocardial slices (Fig.6E). The generation and establishment of the myocardial slices model was performed as previouslypublished (Watson, S. et al. Preparation of viable adult ventricular myocardial slices from largeand small mammals. Nat Protoc 12, 2623–2639 (2017). https: / / doi.org / 10.1038 / nprot.2017.139). Briefly, the myocardial tissue was collected in the operation theatre directly after resection from the patients heart and immediately placed into ice cold KCl-solution to inhibit contraction and metabolic stress. The tissue pieces were transferred to the vibratome and sectioned into 100 μm S13211WO / Universität Heidelberg thick slices during continuous perfusion and oxygen insufflation into the buffer. After cutting into rectangular shapes, the slices were clamped up onto elastic frames aligned by the myocardial fibre direction and placed into 12-well plated filled up with culture media and placed into an electrical stimulation station, where the heart tissue is paced continuously (Fig.6F). HomologousT-cells from the same patient (100.000 – 800.000 per slice) and the anti-CD3-anti-LTBP2bispecific antibody construct (No.: 15452.1 at 5 or 20nM) or equal volumes of PBS were added into the culture dish and incubated for 24 or 48h prior to embedding into TissueTek for cryo- sectioning and immunofluorescence staining (Fig.6G). In human fibrotic hearts with high deposition of LTBP2 a treatment “ex-vivo” with the T-cellengaging anti-LTBP2 bispecific antibody increased the relative amount and area of apoptotic cellsafter 24h (Fig.6H). Functional assays measuring potential functional improvements by changes in contractility will be performed to elucidate a potential therapeutic option for patients. Murine anti-LTBP2 bispecific constructs: To validate anti-LTBP2 bsMAb effects in a murine model in vivo, three different murine constructs were generated characterized by the following protein sequences and structural designs. All of them bind to murine LTBP2 and the first two have a quadruple antibody structure and do also bind to CD3 (No.: 15525.1, Fig.7A) on T-cells or CD16 (No.: 15524.3, Fig.7B) on natural-killer cellsand other Fc-receptor expressing immune cells, respectively. The last is on represents acostimulatory anti-LTBP2-anti-mCD28 bispecific antibody (No.: 15902.2, Fig. 7C) targeting T- cells. The same LTBP2 target-specific sequence was used as for the previous murine CAR-T cell generation.We validated the above noted constructs in vitro and in vivo to elucidate the function, killingefficiency and also systemic effects on cardiac function after myocardial infarction. Unless stated differently, the same methods were used as described in the initial patent description. First, we performed killing-assays using the xCELLigence (Agilent) platform, which enables a real-time measurement of cell viability by calculating a cell-index from impendence changes on theplate bottom. We seeded 30.000 primary murine cardiac fibroblasts per well (C57BL / 6N) into a gold microelectrode 96-well plate (E-plate, Agilent) and allowed them to settle down and stabilize for 20 hours within the platform placed into a cell culture incubator at 38°C and 5% CO2. In the meantime, murine T or NK cells were isolated from murine (C57BL / 6N) splenocytes using MACSmicrobead kits for untouched T or NK cell isolation (Miltenyi Biotec) and T-cells were stimulatedfor 24h with interleukin-2 and kept in culture using respective T or NK cell media.100.000 T-cells or 60.000 NK cells were pipetted into the wells following the addition of the anti-CD3 / anti-LTBP2 (No.: 15525.1, Fig.5) or anti-CD16 / anti-LTBP2 (No.: 15524.3, Fig.6), constructs at 10 or 100nM concentrations or PBS to T- / NK cell control (Fig.8A-B). The killing effects were monitored for 24hafter antibody exposure by continuous real-time cell-index measurement. Cell-index wasnormalized to fibroblast-only controls to express relative cell viability. All conditions were run in triplicates and mean+ / - standard deviation was calculated (dashed lines) (Fig.8A-B). Both bsMAb constructs revealed a very potent and fast killing of LTBP2-expressing murine fibroblasts in the presence of the respective T- or NK cells. S13211WO / Universität Heidelberg After in vitro validation of the killing properties and functional characterization, we applied the bsMAbs for in vivo treatment of post-myocardial infarction remodelling in mice. Ischemia- reperfusion injury was performed as described by ultrasound-guided minimal-invasive transient occlusion of the left coronary artery for 45 min in female 10-week old C57BL / 6N mice. At day 1after infarct, cardiac troponin T and left ventricular ejection fraction (LVEF) were measured toquantify infarct size for equal randomization into the different groups (Fig. 8C). At day 4 the injection of 0.5mg / kg body weight bsMAb or control antibody was performed via the tail veins, followed by echocardiographic assessment of the cardiac function at day 7, 14 and 28 after infarct. The treatment with both murine constructs, the anti-CD3 / anti-LTBP2 (No.: 15525.1) or anti-CD16 / anti-LTBP2 (No.: 15524.3) lead to a significant improvement in several parameters ofcardiac function, such as LVEF (Fig.8D-E). The strongest effects were seen at day 28 (Fig.8E). The body weight remained unchanged (Fig.8F) and no adverse side effects were observed. The anti-CD3 / anti-LTBP2 induced a beneficial remodelling of the heart resulting in significantly lower heart weight (Fig.8G), consistent after normalization to tibia length (Fig.8H). The improved andreduced dilatation of the heart was also documented by echocardiographic volume and areameasurements of the left ventricle in parasternal long-axis B-mode traces (Fig.8I-K). Thus, anti- LTBP2 bsMAb constructs improve cardiac function following ischemia-reperfusion injury and might represent an effective therapeutic option with high translation potential for human heart failure development after cardiac injury. Functional relevance of LTBP2 in the context of myocardial ischemia reperfusion injury In order to investigate the functional relevance of LTBP2 in myocardial ischemia reperfusion injury (IR), we leveraged a constitutive LTBP2 knockout (KO) mice. LTBP2-KO mice were generated inthe laboratory of Robert Mecham, as described previously (Bodmer et al., DevelopmentalDynamics, 2023, https: / / doi.org / 10.1002 / dvdy.651). Both LTBP2-KO and wild-type (WT) control mice (C57BL / 6J background) were subjected to 60 minutes of echocardiography-guided IR injury to induce myocardial infarction (MI). Cardiac function was systematically assessed using echocardiography at days 1, 7, 14, and 28 post-IR (Fig. 9A). On day 28, mice wereeuthanized, and their hearts and other organs were collected for histological and molecularanalyses. Representative echocardiographic images illustrate changes in left ventricular morphology and function over time (Fig.9B). Images from the same WT mouse at day 1 (upper left) and day 28(lower left), alongside images from a LTBP2-KO mouse at day 1 (upper right) and day 28 (lowerright), demonstrate progressive differences in cardiac remodelling between both groups. While infarct size has been similar between WT and KO mice on day 1, LTBP2-KO mice exhibited a marked reduction in infarct size compared to WT controls (Fig.9C). Additionally, the assessment and quantification of multiple echocardiographic parameters, such as left ventricular ejectionfraction (LVEF), global longitudinal strain (GLS) and end-systolic volume (ESV) show animprovement of cardiac function over time in LTBP2-KO mice, suggesting attenuated cardiac remodelling and enhanced myocardial repair in the absence of LTBP2 (Fig.9D). S13211WO / Universität Heidelberg To further validate changes in fibrosis, we obtained cryosections at multiple planes (1 mm in between each plane) throughout the left ventricle. We obtained 4 different planes per heart which were stained using Picrosirius Red to visualize collagen deposition. After imaging using a Zeiss Axioscan Z1, downstream analysis and quantification were performed using Qupath software.Representative images show fibrotic regions across planes of the left ventricle, comparing WThearts (Fig. 9 E, upper row) with LTBP2-KO hearts (Fig. 9E, lower row). Quantification of the fibrotic area revealed that LTBP2-KO mice exhibited significantly reduced cardiac fibrosis compared to WT controls (Fig.9F). These findings suggest a protective effect of LTBP2 deficiency against post-ischemic fibrosis and remodelling. Examining fibroblast activation at day 28, we assessed the expression of Periostin (POSTN), a well-established marker of activated cardiac fibroblasts. Immunofluorescence staining was conducted using an anti-POSTN antibody (Abcam, ab215199) following the initially described protocol. Heart sections collected at day 28 post-IR were stained for POSTN. Representativeoverview images (upper row) and high-magnification images of fibrotic areas (lower row) revealedsignificantly lower POSTN expression in LTBP2-KO hearts compared to WT controls (Fig.9G). Quantification of the POSTN-positive area (Fig. 9H) further confirmed a marked reduction in POSTN expression on the protein level in LTBP2-KO mice. These findings suggest that the absence of LTBP2 limits fibroblast-mediated extracellular matrix remodelling, thereby reducingfibrotic scarring following cardiac injury.Functional relevance of LTBP2 in the context of liver fibrosis Investigating liver fibrosis in a mouse model using carbon tetrachloride (CCl₄)-induced injury, weutilized LTBP2-KO mice to assess the role of LTBP2 in modulating liver fibrosis. Both LTBP2-KOand WT control mice (14–16 weeks old, male and female) received CCl₄ injections intraperitoneally (2.5 µl / g body weight, 25% CCl₄ in corn oil) three times per week for four weeks (Fig. 10A). On day 28, mice were euthanized, and their liver and other organs were collected for histological and molecular analyses. Body weight was monitored weekly, with WT mice showing significant weight loss upon day 7, while LTBP2-KO mice maintained stable body weight throughout the study (Fig.10B). Cytotoxicity of CCl₄ was assessed by measuring serum levels of aspartate transaminase and alanine transaminase at day 28, show no significant difference between groups, indicating similarhepatotoxic effects of CCl₄ in both groups (Fig. 10C-D). For each mouse, parts of the medial, leftand right liver lobes were snap frozen for histology. The PSR staining was performed using the same protocol as described above. Representative PSR staining images at day 28 revealed fibrotic areas (dark grey) in liver sections from both groups (Fig.10 E). Downstream analysis and quantification of the fibrotic areas (PSR positive) in percentage across the whole section wasperformed within Qupath. LTBP2-KO mice exhibited significantly reduced levels of liver fibrosiscompared to WT controls, suggesting that LTBP2 plays a role in promoting fibrosis in this model (Fig.10F). To investigate the immune-fibroblast crosstalk, immunofluorescence staining using an anti-CD68 antibody (Biolegend, clone FA-11) and DAPI was performed to visualize macrophage infiltration. Representative images show DAPI counterstain alongside CD68 staining in both S13211WO / Universität Heidelberg groups (Fig. 10G). CD68+cells were annotated in sections from both groups and a machine learning algorithm was generated in Qupath to count CD68 positive cells in all sections. WT mice exhibited higher CD68⁺ cell infiltration in the fibrotic liver, whereas LTBP2-KO mice showed significantly reduced macrophage infiltration (Fig. 10H). These results indicate a functionalrelevance of LTBP2 in the formation of fibrosis and immune cell infiltration, modulating the diseaseprogression in liver fibrosis.

[0003] S13211WO / Universität Heidelberg References: 1. Amrute, J. M., Luo, X., Penna, V., Bredemeyer, A., Yamawaki, T., Heo, G. S., ... &Lavine, K. (2022). Targeting the Immune-Fibrosis Axis in Myocardial Infarction and Heart Failure. bioRxiv, 2022-10. https: / / doi.org / 10.1101 / 2022.10.17.512579 2. Sicklinger, F., Zhang, Y., Lavine, K. J., Simon, N., Bucher, V., Jugold, M., ... &Leuschner, F. (2020). A minimal-invasive approach for standardized induction of myocardial infarction in mice. Circulation research, 127(9), 1214-1216.https: / / doi.org / 10.1161 / CIRCRESAHA.120.317794 3. Barde, Isabelle, Patrick Salmon, and Didier Trono. "Production and titration oflentiviral vectors." Current protocols in neuroscience 53.1 (2010): 4-21. doi.org / 10.1002 / 0471142301.ns0421s53 4. Matthias Bozza et al. ,A nonviral, nonintegrating DNA nanovector platform for thesafe, rapid, and persistent manufacture of recombinant T cells. Sci. Adv.7,eabf1333 (2021). DOI:10.1126 / sciadv.abf1333

Claims

S13211WO / Universität Heidelberg Claims Claim 1: Molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2)comprising at least one LTBP2 binding domain binding to membrane-associated LTBP2for use in treatment or prevention of a disease associated with LTBP2 expressing fibroblasts, wherein the disease associated with LTBP2 expressing fibroblasts is fibrosis. Claim 2: Molecule targeting membrane-associated LTBP2 according to claim 1, wherein LTBP2is associated to the membrane in the LTBP2 expressing fibroblasts.Claim 3: Molecule targeting membrane associated LTBP2 for use according to claim 1 or 2, wherein the molecule targeting membrane associated LTBP2 is selected from the group consisting of a chimeric antigen receptor (CAR), an antibody, a bispecific antibody, a bispecificantibody linking T cells with target cells (a bispecific T cell engager), a single domain antibody(nanobody), an antibody-drug conjugate and a small molecule. Claim 4: Molecule targeting membrane-associated LTBP2 for use according to any one of the preceding claims, wherein the molecule targeting membrane associated LTBP2 is selected fromthe group consisting of a cell expressing a CAR, an antibody linking immune cells and LTBP2expressing fibroblasts. Claim 5: Molecule targeting membrane-associated LTBP2 for use according to any one of the preceding claims, wherein the LTB2 binding domain binds an LTBP2 epitope that allows to forman immune synapse between the LTBP2 expressing fibroblasts and an immune cell to mediatekilling of LTBP2 expressing fibroblasts. Claim 6: Molecule targeting membrane-associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain comprises a variable lightchain and a variable heavy chain forming a binding site capable of recognizing membraneassociated LTBP2. Claim 7: Molecule targeting membrane-associated LTBP2 for use according to any one of the preceding claims, wherein the LTBP2 binding domain is a single chain variable fragment (scFv)or a Fab fragment or a single domain Antibody (sdAb).Claim 8: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain binding to membrane- associated LTBP2comprises a variable heavy chain comprising a CDR1 having the amino acidsequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acid sequence of SEQ ID NO: 6, orS13211WO / Universität Heidelberg wherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2 comprises a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ ID NO: 35, a variable light chain comprising a CDR1 having theamino acid sequence of SEQ ID NO: 36, a CDR2 having the amino acid sequence LVS andCDR3 having the amino acid sequence of SEQ ID NO:

38. Claim 9: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2comprises a variable heavy chain having a sequence identical to or at least80% identical to the sequence set out in SEQ ID NO:

7. Claim 10: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2 comprises a variable light chain having a sequence identical to or at least80% identical to the sequence set out in SEQ ID NO:

8. Claim 11: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2 comprises a single chain variable fragment (scFv) comprising a variableheavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence of SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 and CDR3 having the amino acidsequence of SEQ ID NO: 6, orwherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2 comprises a single chain variable fragment (scFv) comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ ID NO: 35, a variable light chain comprising aCDR1 having the amino acid sequence of SEQ ID NO: 36, a CDR2 having the amino acidsequence LVS and CDR3 having the amino acid sequence of SEQ ID NO:

38. Claim 12: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the at least one LTBP2 binding domain binding to membrane-associated LTBP2 comprises a variable heavy chain having a sequence identical to or at least80% identical to the sequence set out in SEQ ID NO:

7. Claim 13: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, at least one LTBP2 binding domain binding to membrane-associated LTBP2comprises a single chain variable fragment (scFv) comprising a variable light chain having asequence identical to or at least 80% identical to the sequence set out in SEQ ID NO: 8.S13211WO / Universität Heidelberg Claim 14: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the molecule targeting membrane associated LTBP2 is a CAR comprising from N-terminus to C-terminus: -a scFv capable of biding to membrane associated LTBP2,- optionally at least one co-stimulatory domain, and- an activating domain.Claim 15: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the molecule targeting membrane associated LTBP2 is a CARcomprising from N-terminus to C-terminus:- the scFv as defined in claims 11 to 13,- optionally at least one co-stimulatory domain, and- an activating domainClaim 16: Molecule targeting membrane associated LTBP2 for use according to claim 14 or 15,wherein the at least one co-stimulatory domain is selected from the group consisting of CD28 and 4-1BB. Claim 17: Molecule targeting membrane associated LTBP2 for use according to claim 14 to 16,wherein the activating domain is CD3zeta.Claim 18: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the molecule targeting membrane bound LTBP2 is a CAR comprising from N-terminus to C-terminus: -the scFv as defined in claims 11 to 13,- a CD28 co-stimulatory domain,- a 4-1BB co-stimulatory domain, and- a CD3zeta activating domain.Claim 19: Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding claims, wherein the CD28 co-stimulatory domain has a sequence identical to or at least 80% identical to SEQ ID NO:

11. Claim 20: Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding claims, wherein the 4-1BB co-stimulatory domain has a sequence identical to or atleast 80% identical to SEQ ID NO:

12. Claim 21: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the CD3zeta activating domain has a sequence which is identical toor at least 80% identical to SEQ ID NO: 13.Claim 22: Molecule targeting membrane associated LTBP2 for use according to any one of claims 1 to 13, wherein the molecule further comprises at least one engineered immune cell binding domain binding to an immune cell.S13211WO / Universität Heidelberg Claim 23: Molecule targeting membrane associated LTBP2 for use according to any one of claims 1 to 13 and 22 wherein antigen binding molecule is a multispecific antigen binding.Claim 24: Molecule targeting membrane associated LTBP2 for use according to claims 22 or 23,wherein the molecule is an antibody. Claim 25: Molecule targeting membrane associated LTBP2 for use according to any one of claims 22 to 24, wherein the molecule is multispecific antibody. Claim 26: Molecule targeting membrane associated LTBP2 for use according to any one of claims 22 to 25, wherein the molecule is a bispecific antibody. Claim 27: Molecule targeting membrane-associated LTBP2 for use according to any one ofclaims 22 to 26, wherein the at least one immune cell binding domain comprises a variable lightchain and a variable heavy chain forming a binding site capable of recognizing an immune cell. Claim 28: Molecule targeting membrane-associated LTBP2 for use according to any one of claims 22 to 27, wherein the immune cell binding domain is a single chain variable fragment(scFv) or a Fab fragment or a single domain Antibody (sdAb).Claim 29: Molecule targeting membrane-associated LTBP2 for use according to any one of claims 22 to 28, wherein the at least one immune cell binding domain binding to an immune cell comprises at least one domain selected from the group consisting of CD3 binding domain,CD28 and CD16 binding domain.Claim 30: Molecule targeting membrane-associated LTBP2 for use according to any one of claims 22 to 29, wherein the at least one immune cell binding domain binding to an immune cell comprises an CD3 binding domain, preferably an CD3ε binding domain. Claim 31: Molecule targeting membrane-associated LTBP2 for use according to any one of claims 22 to 30, wherein the antigen binding molecule shows an increased binding to an immune cell compared to a wild type LTBP2 antibody.Claim 32: Molecule targeting membrane-associated LTBP2 for use according to any one ofclaims 22 to 31, wherein the molecule is capable of linking a target cell expressing membrane associated LTBP2 with the immune cell. Claim 33: Molecule targeting membrane-associated LTBP2 for use according to any one ofclaims 22 to 31, wherein the LTBP2 binding domain binds an LTBP2 epitope that allows to forman immune synapse between the CAR T cell to mediate killing of LTBP2 expressing fibroblasts. Claim 34: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein the disease associated with LTBP2 expressing fibroblasts is selectedS13211WO / Universität Heidelberg from the group consisting of fibrosis in the context of the cardiovascular disease, kidney fibrosis, lung fibrosis and liver fibrosis. Claim 35: Molecule targeting membrane associated LTBP2 for use according to claim 15,wherein fibrosis in the context of the cardiovascular disease is selected from the groupconsisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiac fibrosis. Claim 36: Molecule targeting membrane associated LTBP2 for use according to any one of thepreceding claims, wherein at least one echocardiographic parameter is improved.Claim 37: Molecule targeting membrane associated LTBP2 for use according to any one of the preceding claims, wherein at least one echocardiographic parameter is improved after myocardial infarction. Claim 38: Molecule targeting membrane associated LTBP2 for use according to any one of claim 36 or 37, wherein at least one echocardiographic parameter is selected from the group consisting of ejection fraction, fractional area change and end-diastolic volume.Claim 39: Molecule targeting membrane associated LTBP2 for use according to claim 38,wherein the ejection fraction is left ventricular ejection fraction and / or right ventricular ejection fraction. Claim 40: Molecule targeting membrane associated LTBP2 for use according to claim 38 or 39,wherein the end-diastolic volume is left ventricular end-diastolic volume and / or right ventricularend-diastolic volume. Claim 41: CAR targeting LTBP2, comprising: -comprising at least one LTBP2 binding domain binding to membrane associatedLTBP2,- optionally at least one co-stimulatory domain, and- an activating domain.Claim 42: CAR according to claim 41, wherein the LTBP2 binding domain binding to membraneassociated LTBP2 is a scFv.Claim 43: CAR according to any one of claims 41 and 42, wherein the CAR is capable of eliciting cell-mediated killing of the LTBP2 expressing fibroblasts.Claim 44: CAR according to any one of claims 41 to 43, wherein the LTB2 binding domain bindsan LTBP2 epitope enabling the CAR to form an immune synapse between the CAR T cell to mediate killing of LTBP2 expressing fibroblasts.S13211WO / Universität Heidelberg Claim 45: CAR according to any one of claims 41 to 44,wherein the CAR comprises a LTBP2 binding domain comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having theamino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO:5 and CDR3 having the amino acid sequence of SEQ ID NO: 6, or wherein the CAR comprises a LTBP2 binding domain comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ IDNO: 35, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ IDNO: 36, a CDR2 having the amino acid sequence LVS and CDR3 having the amino acid sequence of SEQ ID NO:

38. .Claim 46: CAR according to any one of claims 41 to 45, wherein the CAR comprises a LTBP2binding domain comprising a variable heavy chain having a sequence identical to or at least 80% identical to the sequence set out in SEQ ID NO:

7. Claim 47: CAR according to any one of claims 41 to 46,wherein the CAR comprises a LTBP2binding domain comprising a variable light chain having a sequence identical to or at least 80%identical to the sequence set out in SEQ ID NO:

8. Claim 48: CAR according to any one of claims 41 to 47,wherein the at least one co-stimulatory domain is selected from the group consisting of CD28 and 4-1BB. Claim 49: CAR according to any one of claims 41 to 48,wherein the activating domain is CD3zeta. Claim 50: CAR according to any one of claims 41 to 49,wherein the molecule targetingmembrane associated LTBP2 is a CAR comprising from N-terminus to C-terminus:- the scFv as defined in claims 4 to 6,- CD28 co-stimulatory domain,- 4-1BB co-stimulatory domain, and- CD3zeta activating domain.Claim 51: CAR to any one of claims 48 to 50, wherein the CD28 co-stimulatory domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO:

11. Claim 52: CAR according to any one of the claims 48 to 51, wherein the 4-1BB co-stimulatorydomain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO:12.S13211WO / Universität Heidelberg Claim 53: CAR according to any one of the claims 48 to 52, wherein the CD3zeta activating domain has a sequence which has a sequence which is at least 80% identical to SEQ ID NO: 13.Claim 54: Nucleic acid encoding the CAR according to any one of claims 41 to 53.Claim 55: Vector comprising the nucleic acid according to claim 54. Claim 56: Cell modified to express the CAR according to any one of claims 22 to 28. Claim 57: Cell according to claim 56, wherein the cell is a leukocyte, or a myeloid cell. Claim 58: Cell according to claim 57, wherein the leukocyte is a lymphocyte, wherein the lymphocytes is preferably a T cell or NK cell. Claim 59: Cell according to claim 58, wherein the myeloid cell is a macrophage or a monocyte. Claim 60: Molecule targeting membrane associated LTBP2,comprising -at least one LTBP2 binding domain binding to membrane associated LTBP2 and- at least one engineered immune cell binding domain binding to an immune cell.Claim 61: Molecule targeting membrane associated LTBP2 according to claim 60, wherein the molecule is a multispecific antigen binding molecule.Claim 62: Molecule targeting membrane associated LTBP2 according to claim 61, wherein themolecule is an antibody. Claim 63: Molecule targeting membrane associated LTBP2 according to claim 63, wherein the antibody is a multispecific antibody. Claim 64: Molecule targeting membrane associated LTBP2 according to claim 63, wherein the multispecific antibody is a bispecific antibody. Claim 65: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to64, wherein the LTBP2 binding domain is a single chain variable fragment (scFv) or a Fabfragment or a single domain Antibody (sdAb). Claim 66: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 65, wherein the immune cell binding domain is a single chain variable fragment (scFv) or a Fabfragment or a single domain Antibody (sdAb).Claim 67: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 66, wherein the at least one LTBP2 binding domain binding to membrane associated LTBP2S13211WO / Universität Heidelberg comprises a variable light chain and a variable heavy chain forming a binding site capable of recognizing membrane associated LTBP2. Claim 68: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to67, wherein the at least one immune cell binding domain binding to an immune cell comprises avariable light chain and a variable heavy chain forming a binding site capable of recognizing an immune cell. Claim 69: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to68, wherein the molecule targeting membrane associated LTBP2 comprises a LTBP2 bindingdomain comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 1, a CDR2 having the amino acid sequence of SEQ ID NO: 2, a CDR3 having the amino acid sequence SAN, a variable light chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 4, a CDR2 having the amino acid sequence of SEQ ID NO: 5 andCDR3 having the amino acid sequence of SEQ ID NO: 6, orwherein the molecule targeting membrane associated LTBP2 comprises a LTBP2 binding domain comprising a variable heavy chain comprising a CDR1 having the amino acid sequence of SEQ ID NO: 33, a CDR2 having the amino acid sequence of SEQ ID NO: 34, a CDR3 having the amino acid sequence of SEQ ID NO: 35, a variable light chain comprising a CDR1 havingthe amino acid sequence of SEQ ID NO: 36, a CDR2 having the amino acid sequence LVS andCDR3 having the amino acid sequence of SEQ ID NO:

38. Claim 70: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 69, wherein the molecule targeting membrane associated LTBP2 comprises a LTBP2 bindingdomain comprising a variable heavy chain having a sequence identical to or at least 80%identical to the sequence set out in SEQ ID NO:

7. Claim 71: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 70, wherein the at least one immune cell binding domain binding to an immune cell comprisesat least one domain selected from the group consisting of CD3 binding domain, CD28 andCD16 binding domain. Claim 72: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 71, wherein the at least one immune cell binding domain binding to an immune cell comprisesan CD3 binding domain, preferably an CD3ε binding domain.Claim 73 Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 72, wherein the molecule is capable of linking a target cell expressing membrane associated LTBP2 with the immune cell. Claim 74: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 73, wherein the LTBP2 binding domain binds an LTBP2 epitope that allows to form an immune synapse between the LTBP2 expressing fibroblasts and the immune cell to mediate killing of LTBP2 expressing fibroblasts.S13211WO / Universität Heidelberg Claim 75: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 74, wherein the antigen binding molecule shows an increased binding to an immune cell compared to a wildtype LTBP2 antibody. Claim 76: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to 75, wherein the at least one immune cell binding domain binding to an immune cell is engineered show an increased binding to an immune cell compared to an wt LTBP2 antibody.Claim 77: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to74, wherein the molecule is capable of linking a target cell expressing membrane associated LTBP2 with the immune cell. Claim 78: Molecule targeting membrane-associated LTBP2 according to any one of claims 60 to77, wherein the immune cell binding domain binding comprises mutations in the Fc domain thatenhance the interaction of CD16 compared to a wt Fc domain. Claim 79: Nucleic acid encoding the molecule targeting membrane associated LTBP2 according to any one of claims 60 to 78. Claim 80: Vector comprising the nucleic acid according to claim 79. Claim 81: Nucleic acid according to claim 54 or 79, or vector according to claim 55 or 80, for use in treatment or prevention of a disease associated with LTBP2 expressing fibroblasts,wherein the disease associated with LTBP2 expressing fibroblasts is fibrosis.Claim 82: Nucleic acid according to claim 81 or vector according to claim 81, for use according to any one of the preceding claims, wherein the disease associated with LTBP2 expressing fibroblasts is selected from the group consisting of fibrosis in the context of the cardiovasculardisease, kidney fibrosis, lung fibrosis and liver fibrosis.Claim 83: Nucleic acid according to claim 82 or vector according to claim 82, wherein fibrosis in the context of the cardiovascular disease is selected from the group consisting of fibrosis in the context chronic heart failure, fibrosis in the context myocardial infarction and cardiac fibrosis. Claim 84: Use of molecule targeting membrane associated latent-transforming growth factor beta-binding protein 2 (LTBP2) in diagnosing a disease associated with LTBP2 expressing fibroblasts, fibrosis, chronic heart failure, and / or myocardial infarction.

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