HU230186B1 - Preparation for delivery of agents to solid tumours - Google Patents

Description

DISCLOSURE OF THE INVENTION

The present invention relates to genetically modified rnesenchymal stem cells (mesenehymal stem cells, fe5) and MSCs. Up to said cars or allogeneic MSCs, the expression of galactin-a is silenced with galecin-1-specific siRNA. In particular, M8Cs can be used as mediators for the digestion of gslectin-1, expressed in the five Moors, and can be modified to produce tumor suppressants, in particular galecal inhibitors. Bsirrjekiálí the MSC ~ k eliávolkhaiók transzgetókus a suicide gene 'using the application, which will destroy the therapeutic IO MSCs used as a carrier after _therapy: One preferred. This invention provides the possibility of local treatment of solid tumors, thus reducing the risk of side effects.

TECHNICAL STATE

A. meseneisymal: stem cells or stromal cells (MSCs) are non-lipolytic progenitor cells. meKsA field unan-- s / .acuA seujo \ c, pousegte / studs e- «.sortvelu 'kotes / ovcbe ^ epc

IS has many oesophageal secretions, some produce maturation and division of hematopoietic stem cells (Valiién et al, 20O8), is most often isolated from bone marrow (CsV), but can be extracted from many fetal and adult tissues (Pítteoger et al, 1999 )

One of the horses of the MSCs is to help and regulate the differentiation of blood cells. in bone marrow (Plttenger et al, 1999; Chiunberlaln et ah, 2007). In addition, MSCs are well characterized by their strong 1 «waiting and um? The suppressive effect of v / vo lumum suppressive effect on virtually all cells of the bone system These effects are exerted, inter alia, by the 1 -.eh polyolsasa.td ¢ .Bartbokuneu et al 2OO2, Os \ -, ο Α G al 2uf, allogeneic cells and milogens. > 2 ', al ~> etuk tab'. 'Xépte majesty (Gfennie et al, 2005) or apoptosis. (Plumas et al., 2005) by inducing alteration of cytoclonal production (Aggarwal and Eifcugcr 1815-22) and inhibiting the maturation of anti-demetic cells and antigenic culture (T'feyth et al., 2005; Hanta et al., 2006). MSCs have also been shown to prolong the survival of gonadal transplants (Batíholomew ski al, 2092) and to reduce experimental transparency in model-induced sefer phosphate multiplex (Zappia et al, 2005; Gerdon et al., 2007), collagen inducer arth, 2007). and type 1 diabetes (Lee et al, 200b; L'rbau et al, 20O8). More importantly, the infusion of M8C-h has been shown to have a beneficial effect in the treatment of steroid-refractory gralt-versss-hosiery in patients with allogeneic CSV transplantation (Rmgáén et al, 2006).

As described above, MS € controls the immune response by inhibiting the function of Th1, ohotoxic T cells, and NK cells (Gordoni, E. et al., 200: Alma J. Hanta et al., 2007). they are involved in the regeneration of damaged tissues but also have a potent anti-inflammatory effect.

Although the exact mechanism of these effects of MSCs remains to be elucidated, direct cell-cell interaction and soluble fecal trees have been reported in the literature.

3040a 5-34Ö5A / SG / L2s

Soluble Factors Produced by MbC,? -Polylet Growth Factor, Trans-Spermal Growth Factor-β (TGF-β; Di Nieola et al., 2002; prosiaglandin-2 C'PGE-2; Aggarwal and Pktenger, 205), mdolamism- X6-dioxygenase (ISO: Meise.het ah 2004), nitric oxide (NO; Bato etak 200 / Ken et al, 20). synbil HLA-G (Selmám et al, 2008} and IL10 - all of them are anti-inflammatory mediators (Fáye Ii. Chen, 2008). Conversation between MSGs and immune cells ultimately also affects the anti-inflammatory function of MSCs: activated T-cells and n-NR cells produce inflammatory euthytes, I.FN-γ and TNF, which then act on the bed to effect MSCs, which are the major inhibitors of the PGF.2-L γ-F class. -y gzen surface also stimulates 1D6 expression / expression of MSCs, resulting in tryptophan deficiency

10: Growth occurs by inhibiting T and NR divisions (XM.Ryan. 2007; R, English 2007). However, the data do not prove that any of the phosphorus produced by MSCs is responsible in itself for the presence of mimicry soap effect.

As demonstrated in injured or tumorous state dollekbe, externally delivered MSCs migrate to wounds and injured tissues and are localized in the never-healing wound (Stndeny et al. 2004). However, MSCs play an has not always been clarified, for example, in an in vivo animal model of Raposi sarcome, intravenous human MSCs migrated into the tumor and significantly inhibited tumor growth (Khakoo et al. 2006). Other studies have shown that bone marrow-derived stem cells supported tumor growth in mice: when MSCs were co-inoculated with tumor cells, they showed stronger tumor growth, more extensive necrosis, and higher transfusion rates than W, ah 2006). The tumor-promoting properties of MSCs can be explained by the effect of these cells on the nnnmosappressfe and thus the inhibition of the tumor specific immune response (Hanta AJ to Fibbe WE et al). This theory is supported by the result that, overall, melanoma can only grow if 25 MSCs are simultaneously inoculated with the densities (Djoead F et al., 2003) · Az.MbCs are wounds (Wu et al., 2007), ischemic areas (Nagaya et al. 2004) and vascularization of tumor tissues (Sun et al 2005), which is a crucial process in wound hernia and fvnto iedodeshen.

There is also intense debate about the future fate of MSCs in tumorous tissues. According to some of these authors, MSC-fc, as mileite-derived cells, a. Lymphocytoma (one of the cellular bases of tumor associated dbroblasts, vasodilectin and smooth muscle) (Oswald et ah, 2004). Other publications, however, argue that: MSCs are not stably localized at these sites but influence tnmorgenesis by producing different vectors (TGF-β, TIME, ILI, PGE2, VEGF) (Roorda et al, 2008), so, mig. The M'SCs have a beneficial effect on bone marrow renal dllfe55, regulation of the immune response, and wound healing, but are also detrimental to tumor growth and metastasis (G. Fazermee 2008). The mechaBizBms behind this effect is yet to be discovered.

A '1Ά Í ntotovo anvitoumm to có / to, t de:

Now »Mto has been used in a few weeks as a delivery device for delivering antitumor agents: tumors. M. Stndersy et al., (M. Studeny et al., 2002) genetically modified:

S was inhibited by the introduction of MSCs & Immun A375SM. melanoma cell growth in immuno-mined mice. Tomous MSCs produced 1ΕΝ-β-άί, an inhibitor of tumor growth. A more detailed and general description was published in WO-033737998 ("Local Produetien and / or Deltaery of Anti-Aging Agents by Fluids, ceti praers").

Later, M, Stadeny et al. an MSC expressing human α-β in an xenograft mouse model was inoculated into human MDA 231 breast carotid or A375SM melanoma tumors. B8C ~ fc, incorporated into the classroom, inhibited growth of mice and increased survival of mice (M Studeny staff 2004).

Similar results were obtained by A. feakamizo and myst., Who found that the human IRG-1EMβ-producing human was increased MSü-fc s / ugudiculously in the gymnastically 1) 87 xeogeneic glial gna cells lyratoamically: survival of mice <A. Nafamizo et ab 20Θ5).

A. In DE 102006020307 and DEI0200020444, letoviral and / or retroviral tomato-like apoptosis-inducing ligand (ITRA1L) similar to lymphatic uveitis factor (THE) have been formulated to provide a stable expression vector, which still retains the same vector the 3 'end of which contains a self-activating revenge terminal repeat sequence. These cells have been successfully used in trarter implant experiments in pseudomodel models and in the treatment of tutors.

No. 102000345720 discloses a system for treating brain tumors using a telSC carrying the vector encoding a suicide gene (Herpes-sunptex virus tbnldin-chitose (HSVtk) machine). This allows: the suicide gene expression gene thereby produces an enzyme that converts a precursor (prodrug) to produce a fermakotegially active umectectase that kills lysnidine kinase transgene producing cells, inhibiting their division. The above system is capable of treating brain tumors by destroying or suppressing tomocytes, and is particularly useful in the treatment of glomerulus,

E. Marit gave an overview of the use of CSV-derived MSCs as cellular mediators (F. Marini, 2006).

8. Standing in an attempt to determine the future direction of MSC refill strategies (B. Ball, 2007),

Among the most recent. Headfemées are L. Kneerova et al. who differentiated MSC from fat cells expressing eysteine deamylase and found that these signals were inhibited by the recombinant cells. Proliferation of xesogenic HT-29 hanthan adenocarcinoma cell line subcutaneously injected with 5-flnorocytosine prodrug (L, Kucerova, 2000),

Recently, transgenic MSCs expressing diabody expressing transgenic MSCs expressing diabody have been implanted with CEA-positive human iron &lt; RTI ID = 0.0 &gt; g: &lt; / RTI &gt; carcinoma (HCT in 16 cells), xml. Human T-cells do this: Transplantation of the follower reduced the growth of TNF. (M. Compfe, 2ÖÖ9),

Gal is a monomeric or homodlmer protein, A homodlmor non-covalently: 2 monomeric Gal-l from bonded subunits are formed by 134 cmin> carbohydrate · binding domain, Gabe has been identified in the following tissues: grown smooth and skeleton, tnnus, lymphatic drainage, prostate, tear, pfecenta, endothelial cells, skin, testis, s / agonies, developing brain, and: retina (Rabinovleh GA 2ÖÖ2, Reriio NE 190k).

Numerous over the past few years; data confirmed the role of Cali-1 in tumor growth in helical secretion, angiogenesis, and tumor nucleation immunoreactivity (Rabinovleh GA 2005}. ). The silence of Gal-1 expression led to the development of a trnorspecific immune response (Rublnstein N et al, 2004, Maiidon V et sl, 2007),

Gal-1 is produced in large quantities in tumor cells to the tumor! Associated connective tissue (skeletal roma) and / or endothelial cells, Therefore, Galta is considered to be a molecular marker of cancer (Camtó et al 2006), Gal-1 is also expressed by MSCs (Katin I et al 2005; Panepueei R et al 2004), but prior art has not determined its intracellular localization (i.e., intracellular secretion or secretion), the role of lileve: fenccphenal in the immunosuppressive activity expressed by MSC-fc and in the induction of T cell apoptosis. The present inventors have surprisingly found that Galeknu's superior blood (Gal-I) is a blood-cell mediated by T-cell apoptosis in the RISC media and thereby contributes to the immunosuppressive environment in tumors.

To the best of the inventors' knowledge, no written or communicated information has suggested, suggested or suggested that genetically modified M5Cs could be cellular mediators of Gal-1 inhibitors. The present invention claims that recombinant MSCs are particularly painful for this purpose. BRIEF DESCRIPTION OF THE INVENTION

The subject of the invention is a recombinant genetically modified (recombinant) snesenepheymal stem cell (MSC) that has been modified to be unable to express and / or secrete gafektln (Gal-1) in the MSC. inhibited (knocked ont) or highly inhibited (knocked out knewed) Preferably, the Gal-f expression is silenced in .MSC. In a preferred embodiment of the invention, the preferred method is Gal-1 expression. to reduce the gap in the siRNA set,

The present invention also relates to: genetically modified mesenchymal stem cell (MSC), which is modified to leave galectin-1 (Gal) for production and / or secretion. and its malignancy is for use in cell therapy of solid tumors.

In a preferred embodiment, the genetically modified M8C is capable of producing another recombinant polypeptide with a compound, preferably a &quot; moretew &quot; activity. As a result,. on the other Tekes: a binary compound introduced into the solid tumor cells or into the lymphatic fluid surrounding the tumor.

very preferably, the additional compound is a GsM blocker, more preferably, the Gabi inhibitor Gabi5 blocking agent, which is as follows. a Gabi derivative, a neu-lysing anti-Gal-1 antibody, its derivatives and a Gabi binding fragment; ollgopepbd atom mimetics, oligopepdd glycemic mimetics, gab inhibitory oligopeptides and polypeptides,

The galectin to be inhibited may be produced by tumor cells, endothelial cells, tumor cells, or endogenous MSCs in the tumor.

In a preferred embodiment, the galectin-1 inhibitor is selected from:

angínex or a derivative thereof capable of binding to γ-1-h, a non-biologically active horseradish of galefetin-1, which fishes as a bait or is inhibited by endogenous Gal-I

In a preferred embodiment, the. low-grade solid tumors respond to the following: lung cancer, vascular and terminal cancers, squamous skin cancers, such as squamous cell carcinoma of the head and neck. egg-kamma, bladder cancer, glioma, thyroid gland cancer, bladderworm cancer, prostate cancer, kidney cancer, breast cancer. Preferably, the cell therapy may be autologous or allogeneic cell therapy, where it is possible: to administer the therapeutic agent repeatedly,

In a further preferred embodiment of the present invention, the mCombinant genetically modified MSC described herein carries a so-called suicide gene that allows for the inducible removal of this genetically modified MSC, e.g. after the required treatment period. The thymidine kinase gene is particularly suited to this task,

The present invention relates to the use of a genetically modified MSC as described herein in the manufacture of a vehicle for delivering a Gab i ·· inhibitor to a tumor, tumor cells or microenvironment of the tumor.

Preferably, the n Gabi inhibitor is a Gabi inhibitor olgopeptide or polypeptide produced by a genetically modified MSC.

Preferably, the genetically engineered MSC is delivered to, delivered to, or migrates to the tumor environment, and is preferably incorporated into the tumorous tissue,

The present invention relates to a kit for delivering a Gabi inhibitor ohgopepid or polypeptide to a solid tumor, to tumor cells or to the tumor microenvironment, said kit comprising genetically modified MbC and a device:

- introducing a gene encoding a Gabi inhibitor five-peptide or polypeptide into a genetically modified MSC, and / or

- genetically modified MSC. to the tumor or to the microenvironment of the tumor.

The present invention further provides a method of treating a patient with solid tumors comprising: Directing horse cell therapy comprising the following steps: administering to a patient genetically modified mesenohymal stem cells (MSCs) which are modified in soda as described above. that they are unable to produce and / or secrete from Gafokin,

- where the recombinant genetically modified MSCs are preferably compatible with M.

- where the genetically modified MSCs are introduced into the tumor microenvironment, they migrate to or from there and integrate into the lumor-MS, thereby competing with endogenous MSCs,

Ga | ~ I ~ can be produced by the Tumor Associated Siphon (TAS) cells.

In a preferred embodiment, the therapeutically detailed therapy is an anthologic or allogeneic cell therapy, comprising the following steps:

- taking samples of klSC contents from a patient or other voluntary donor,

TO - MSC collapse from sample,

- genetic modification in the fonts detailed in at least part of the isolated MSC to reduce Gabi production,

- cell cultivation of so-called genetically modified MSCs.

- therapeutic, genetically modified MSCs. administration to a patient.

Described further is a method for delivering Gal-1 inhibitory eyes to a solid tumor, tumor cell hex or tumor microenvironment, comprising 8 steps of: administering a composition of a genetically modified therapeutic MSC of the present invention to a human or animal subject; MSCs have been modified to the extent that they are not capable of producing Gal-I and / or secretion,

Where genetically modified MBCs are introduced into, or delivered to, the tumor microenvironment. germinate and integrate into the ternary mucosa and thereby compete with the endogenous Msek, they can produce Gai-I on the Tumor Associated Siphon. (TAS) cells.

According to a preferred embodiment: a. method further includes the following steps:

- genetic modification of isolated MSCs to reduce production of Gal-1,

- cultivation of genetically modified MSCs, thereby multiplying,

- administering or returning the propagated MSCs to the patient.

Preferably, the rocombinant modification of MSCs is done using sjR \ $ to reduce OaLl protein expression levels,

The disclosure further provides a method for treating a defined rttmorph patient, which comprises administering to a patient a G-1 inhibitor of the present invention by genetically modified Gal-1 negative MSCs, wherein the galectin-1 inhibitor is genetically modified. its modifiers are produced by MSCs and secreted.

Preferably, the Gal-I inhibitor is capable of inhibiting endogenous MSC and G-1-induced T-cell apoptosis and / or neoanglogenesis and / or transfusion (metastasis) of GN-1 derived from striatal or tumor cells.

Preferably, the Gα1-I inhibitor of the invention is a Gal-1 blocking agent (i.e., an inhibitory chemical that binds to the Gal-1 protein) or a Gabi competitor compound,

THE FIGURES. SHORT DESCRIPTION

1) A) Paraffin-embedded, cellulose-derived brown: MSCs were stained with Gal-1: monoclonal: antibody (niAb) (black), B) Gabi expression in human (bMSC) and mouse (tsMSC) mesenchymal. stem cells in SDS> AGE ^: sample buffered half and 12% SD8 in diatomaceous earth. Purified recombinant Gal-1 was used as a control. Proteins were transferred to a nitrocefluorescence membrane and blotted with antI-Gal-1 mAb followed by anti-mouse Ig-HRF ?, and reacted with the EGE Plus detection system.

Figure 2, Gal-1 is secreted and binds to the surface of hMSC. A) Cells were treated with thiodiglycoside 10 (TDG) or left untreated (black line) and left with aatl-Gal mAb or unstained (gray dashed primordial black dashed line), finally to each sample. Atta-4O-labeled labeled anil mouse Ig was administered and assayed by FÁCS assay, Bj The figure mimics the relative intensity of femresenol in the samples.

Figure X. Gal-11-cell epoptotic from MSC Induced by Jurkat (A and .alpha.), Or Akt115 became T-cell brush (Bj with bMSC (A primed B) or m (mouse). Hours were maintained in a co-operative hour (C): Mh were pretreated with 30mM TDG for 30 min or left untreated (A and C), TSCs stained with .MSC and Boeehst 33342 ( gray), the samples were stained with Gal-1 mAh-primed anti-mouse l: gG-NL577 and Altexm-V-Alexa Fluor fehér-white (white), and analyzed by heating tube microscopy. shows the percentage of apoptotic cells

TDG mlesletehen or telescope, counting at least 1000 cells in each sample. Apoptotic cells are indicated by arrows.

Figure 4, MMSC-s Gal-1-sel siRBS ímMSG '^' and sequencing inMSG RN8 ~ sei (scrmnbled RNA, MMSC ^5raí ') stably írarsszfektáhunk ^ Ö or mixed. Gal-1 expression was checked in each cell line from Western biota using rabbit at-Gal-i (Santa Graz). Wild-type and Gal-1 ~ silenced MSCs were maintained in culture for 1 h with Boechst-stained Jurkat cells. The apepVtsG s - ^ 'bow * Vnu'vnt \ - Vcu 1' .um vSS with deatoaahek (feh <.n \ d'ag'am depicts the degree of ahafeukalt upopyxion of kuionho / o VfeC cells. Wild-type MSC induced apoptosis by 1 µm% and compared to cell death caused by other MSC cell lines. Silencing of Gal-1 expression: decreased the apoptotic effect of MSCs,

3Ö Figure 5. 'MSC · Mediated Needles for Apoptosis: The direct interaction of these cells is critical. b.MSCs and Jurkat cells in co-culture (A and B) _; or tranwell membrane (C and G). After incubation for 16 ohms, Koeehst-stained Jutka cells (light gray) were removed by typhoid and Ánnexln-V-Atto 488 (A and C, white): or anit-Gfe-1 rnAhffierihern Lighís 557 (B, D, gray). stained. Samples were examined using a Buorescross microscope. The apoptotic cells are indicated by (green) arrows, E). The graph represents $% of apoptotic cells, with at least 1 killer cell number,

Figure 6. MSCs increase primary tumor development and promote metastasis, A 3xlU ⁵

C57RI / O mice inoculated with roma melanoma cells at different times measured the size of methane lymphomas (A). <Π breast hstooomával (1o ⁵⁾ grafted BaWc egrekben of: injection rock drill 25 days was determined in the primary tumor (B) and the lung weight (C j and metaszíaíikns nodafctsok number (D) The imnorsejiekei. Both tumorféléífel alone or ITE Inoculated with MSC, subcutaneously.

DETAILED DESCRIPTION OF THE INVENTION

The inventors' Isolated MSCs are useful as transport vehicles for transporting Guhl's anti-γ-1 inhibitor inhibitors to the tumor and for the delivery of tenor microfibers. The inventors have shown that the γ-1 key plays a role in the sep-cell apoptosis induced by the lSCs by sep-cell interaction between MSCs and T-cells. thereby healing the tumors to the immune cell.

The MSCs of the present invention are highly capable of producing Gal-1 production and / or secretion, thereby avoiding the binding of the MSC-produced Gal-1 to the inhibition of the δ-1 inhibitor. And the blackheads. váraifenul found that the GAI-1 silenced MSCs apoptötífcns effect degree as rain and _the t-öal. esendesités not affect the MSC ~ fc migratory ability of tumor. Further, the present invention may inhibit tumor immunosuppressant and promote tumor specific

SS expression of immune response against solid tumors.

As described in the state of the art, MSCs play a role in tissue regeneration and have a potent anti-inflammatory effect by targeting various hnmn cells, T and B cells. natural killer cells (inhibit activation of NKI and professional antigen presenting cells, but activate mummy suppressive T-cell function)

The origin of MSC, the majority of factors affecting immune cells, acts as mediators in solubilized media. In contrast, the present inventors have found that Gat-1, which is a protein of high yields produced by MSCs (Figure 1), although sequenced, is immediately bound to complex sugars on the surface of the producing cells or to glycogorugates. ), and glycosylated proteins of the cMracellular matrix (Figure 2). Indeed, when human: or murine MSC25 is co-cultured with: (co-cultured) Tokai leukemic cells or activated T cells, the surface of the latter cells becomes Gal-1 positive: despite the fact that these cells do not express endogenous γ-1. (Figures TA and B). Ola-1 and also Annexin V-positiv T cells confirm that MSC-derived Gabi is the media for T cell apoptosis. (Figure 3). This result is supported by the observation that removal of cell surface Gal-1 from the surface of MSC by TDG washing decreases the degree of T-cell apoptosis caused by MSC mdu30 (Figures 3A and j). production of MS-1 with Gabi speic siRNA is reduced, T-cell apoptosis induced by Gal-1 expression is dramatically reduced compared to wild type or nmek transfected MSC (Fig. 4), Transwell plate experiments (Ia. Examples) that MSC-derived Gab i-induced T-cell apoptosis requires direct cell-to-cell stone apoptosis (Fig. 5). Physical separation of MSC effector cells and T-cell cells in a common medium prevents T-cell apoptosis and in the event that only the medium is able to flow freely between the isolated cells (Trans-T cells do not become Ga-I positive). the experiments, and the fact that Gal-1 does not appear in the fluid of the MSC tissue, support it. ftpgy Gal-1 is not a soluble but a cellular vector. The human and mouse GaM Tekliac A high degree of amino acid homology between the proteins of -1 (consensus amino acids: 94%, sequence identity: 88%) is of interest.

In some tantors, tumor associated striatal and tenor associated endothelium both produce large amounts of 5 Gal-1. (Oaaguy A et al. 2002; Rnbioovich GÁ and body 2007). Specifically, high levels of Gal-1 have been shown in a wide variety of tenors (see Camhy, ß et al. [Glyeobiology Vet No. 1 shoot pages! J? A-i57B, (2006) and Danguy, A, 1 et al.) G & leetins and eaneet Vol.

1572, pages: 285-293, {2002U

The present invention is applicable without limitation to the treatment of metastatic or non-metastatic tumors of the genus Ensber or non-human: metastatic breast cancer, colorectal carcinoma, squamous cell carcinoma of the head, ovarian carcinoma, cataract. giloma, thyroid cancer, pancreatic cancer, prostate cancer, renal cell cancer, etc.

It is believed from literature and experimental data that tumor cells and / or surrounding stromal cells (TÁS) (supporting tissue or endolome) in tumor tissue produce large amounts of Gal-1. GaM. contribute to tumor immunity of tumors, promote neo-nangiogenesis and cancer cell migration. This is the reason for the starbareghna of talab. mcb inhibits the tumor (0: 1-1) and promotes the development of unmnmala ^ z ranuueMencs.

There is evidence in the literature that MSCs are involved in the formation of the temporalized stroma ki20 (Hali, B et al., BBP (2007) 180: 263-385) and that endogenous MSC-fc also migrate into the tumor microenvironment (borning) and cicatricas are integrated; however, this mechanism is still largely unknown, so that tales of mammalian stem cells are considered as a means of delivery for the potential delivery of tumor somaoma precursors and anticancer agents. [Matns. Stndeny et al. Of the National Caneer Instimte, Vol. 96, No. 21, No. 3, 2004; l lah et ah Intern, 'dame Jonrnnl of

Idematology, Vol. 86, No, I, pages 8 to heat, 2097], d nmsouc / pwffi »A,» Wiv ₍

Mezencis malalis, socks can be from healthy volunteers' bone marrow samples ?: (allogeneic) or liposuction from a number of adipose tissue (allogeneic), MSCs can also be obtained from the bone marrow or fat tissue of the patient being treated, termed anabolic cell therapy,

Proposed by ISCT Infernal (bm / i} ibr Cellnlar Thenapy) t Dominíel et ah, Cytehetapy

20: 6, 8: 315-7), the minimum criteria for defhnabs of MSCs are:

- plastic nickel adherents under standard culture conditions;

- CD44. CD73, CD90, and CD105 are positive but CD34 is CD45, CD14, CD11i or 14LA-DR negative, and

- are able to differentiate into osteo, fat and pnese cells.

A M? C ~ ji .. moc n murine ooze (Tumor cells, tumor associated sarcoma and / or endogenous MSC axd re: breast target to be inhibited)

The inventors have found that .MCCs are gene-modifiable in bed to be fool-proof in cancerous therapy: first, therapeutic MSCs need to inhibit the expression of the Gal 1 gene or Gall protein, which promotes the growth of the primary tumor. metastasis In particular, these Gall silenced M5Cs are capable of delivering Gell's agent to the tumor, as the "self" Gall. m enters competency to the site with therapeutic Gall inhibitors

Consequently, the present invention is not directed to Gat-1 production. are capable modified MSCs. There are many technical methods available for the production of these modular MSCs. For example, a very good method would be to knock out) the MS gene in the MSC. A massk tcnvtoseg of the RNA mtettence of $ mRNA) 3 phases λ Gal 'tcrmekxte in ml

1Ö I know arnsseea by technique Is. It can be inhibited. Specialist dives. suitable methods, which were swept in the following publications: Gajrnbv I and rotsi. (2005), Rubbsfeb et al., (2004), te kereereier et al. (20Ö8) or m WÖÖÖÖ08474 (Publication: 200Ö-H119, Inventors: Oamhy, Isabelle).

Because MSCs migrate to tumor tissues, they are suitable for transfecting genes encoding anticancer agents or cytokines. «Maybe to transport them to the tumor. In fact, since MSCs contribute to the formation of the intramolecular heart, it is possible that genetically modified, non-Gallti but non-cancerous MSCs are effective therapeutic intervals. The following publications disclose methods for producing and maintaining MSCs; Nakamizo, A. et al. (2005); Rail, M and missi. (2002 g Stadeny, M et al., (2804); Hali, 8, and Rítsí, (2007); ÖzawaX et al., (2000)).

Within the scope of the invention, non-Galile MSCs may be further modified by expressing a known Gabi inhibitor protein polypeptide. Examples of such soft peptides, such as tumorellees, are well known in the art.

Allogeneic MSCs must be characterized for tissue compatibility (HL A) before being administered to a patient.

The determination of the unit dose of genetically modified MSCs of the present invention is influenced by a number of factors: a. individual diagnosis, severity of hernia, route of administration, patient age, sex, body temperature. Definition of all these is the duty of a specialist,

The present invention relates to a wall of MSCs that produce Gall inhibitory material migrating to the primary or secondary Tamil area.

The Gal I inhibitor may be any molar / chemical produced by genetically modified MSCs that partially or completely reduce Gall activity. The Gal-1 inhibitor directly exerts its effect, for example, by binding to Gall, or indirectly, e.g. by modoteae Gal-1 bphospholose beta, or by activating another Gall inhibitor. A Gall inhibitor is a Gall inhibitor that binds to Gal-1.

One of ordinary skill in the art will appreciate that many methods are suitable for the preparation of a temsgenic MSC producing an oal-1 inhibitor of the invention.

For example, the MSC slide-produced Gal-1 blocking agent may be a non-lysing antibody or derivative thereof, e.g., Fab fragment: either an oligopeptide lequinemimetic or an oligopeptide eocormimethite or kb compounds / molecules wherein the Gabi blocking agent is capable of Block the effect of Gal-I.

A possible Gal-1 inhibitor peptide ph is Anglnex, which is a planned 33 amino acids! stationary peptide with a potent inhibitory effect on angiogenesis. For the WÖ290612S027A1 sled walomha »[published: 2Ö6] 1-30. inventor: Griffioe® Arja.® \ VJ recombinant Asp33-Anglnex potent peptide. The recombinant Auginnex polypeptide is a. by inhibiting proliferation and migration of endothelial cells, it causes endothelial cell disruption and apoptosis. The Angbex receptor on tumor-associated activated endolytic cells (ml-L Although numerous in vitro experiments have shown binding between Gal-1 and Anglnex as a direct link, '' Hljssen et al., 2000 / did not determine the exact binding site of Arsginex. Gala on a molecule.

my main experiments confirm that the molecular target of Angisrex is Gal-I because it is

70% of Anginnex treatment inhibited tumor growth and 5S% of vascular formation in F9 lake carcinoma 12.91G ') mice, while Gal-1 knoek-ουί (Gal-V) mice did not exhibit anticonvulsant activity (Thijsson et al., 2 ()). ). Brandvdjk et al. In 2006, Anglnex was also shown to be an excellent gene therapy approach for cancer, as slower tumor growth was observed when B16F1 (I transfectant mephenoma transfecting cell) was injected into experimental mice.

Numerous additional Gal-1 binding molecules / chemical proteins are described in the literature:

giscosdal polypeptides linked to one or more sugars, in particular ornamental gums (WO03026494). preferably derivatives of lactose, galactose or irirucose, in particular fructose-D-leucine, lacufufe-L-leucb, diacyl-lsxamethylenediamine, etc.

- asialofototin, a complex glycoprotein derivative of tetra-sialic acid,

- Lialis digital lactoside (TEX5),

- ski RNA, eg: slRNA technique described in WO20005 & 474A2, which is capable of reducing the level of Gab1 in the tumor,

- a Gai-1 binding agent, ph: JP20-721-968A, a neural stem cell growing 30 cells! inhibitors capable of inhibiting the binding of Gal-1 and β-1-btegrm.

An embodiment of the invention is safe cell therapy. For example, following the desired repeated application of our variable confereo-prophylactic strategies described in the fonts below, the transplanted MSCs may be removed from the patient's body at the end of treatment by carrying a so-called suicide gene (ph: thymidm kbase) suitable for transport kpnstllyl is my therapy path

Because topical administration of genetically modified Gal-I secreted therapeutic MSCs (ph: Intravenous Injection.}) Provides topical treatment, it is more advantageous than current systemic therapies, e.g., central nervous system tumors where topical treatment cannot be resolved and / or most of the systemically administered agents are ineffective (eg, do not cross the blood-brain barrier).

In these cases, the present invention provides a unique solution.

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1, Materials and Methods: 5. $ iZÍÍ ??. '.? Z $ ífeÁ

The discs were grown JUTKA I -scju penicillin'dreplomíemnel (Hiu ^II! Es Bridge g mis 1 • giutumiunai (2Nal) and supplemented with 5% sgszölött bosjú szétüonnal (FCS, ancestor, Invitrogen) RPMM64Ö (Gtbco, Invitrogen) with tissue culture medium. A human (hMSC) and mouse (mMSO MSCs penieillin / L streptoínieianek gíutammual <2t »M) and 10% FCS se? maintained D.MEM supplemented medium at present, the sejtvnnalakat 5% CO_> -hun 37 ^o C human and mouse lineages were isolated as described above (Vas et al., 2005).

PerlLr.al s mononuclear cells (P8MC) were obtained from healthy donors by centrifugation of blood Fiooll Gradient and activated with PffA (5 pg / ml) for 72 hours. Medhntbas containing 10% FCS and 20 ng / ml 11, -2 were then grown in the cells. Twenty-four hours before the examinations, 24, -2-ß was removed from the medium.

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In this assay, the following reagents were used: Nitroelulose membrane (Schleieher and Sohuelj). rabbit, antl-egor tag HRP (BAikko), BCl plus detection system (Amersham Bloscien.ee), X-ray film (Medvedn RFBk ÁnnexinV - Alexa Fluor 488 (Mofecniar Probes,

Indüogen). AnnexnA - Ali 4 $ 8 (MvxiA unu mouse IgC-Nmlbernl jghts-55 (RAD SyOemsk Flaoromount-G (Southern: Sfelelt). Tilt Molecular Weight Marker (Fermentas), PhytolmmagglntininM, PlinA (Calbi) , doron Gorporaiion), Fiooll-Paqué ™ Boy (Amersham), TrauswelW Penular System (Corning Costa?}. Gal-1 otonoclonal antibodies (clones 2C1 / 6 and 3C1 / AI) were prepared in our laboratory according to standard protocol. It was obtained from Aidrleh.

The nSUPhRIOR RNA Kit is a product of OligoEngine (Seattle, USA). The AttoáSS PCR Marker Kit is from, 1} '\ A Kiohcieoce (Jena, Germany)

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The cell pellet was suspended in cold FACS paste (PBS supplemented with i% FGS and 1% Naase). For the detection of seplblsin Gal-1, anti-human Gal-I mohlonal elution (clone 2C1AS) was used followed by anti-mouse IgG-Atto 488. After washing, cells were lysed in FACS buffer, to which dead cells were added with propidium kungedide. To trace the membrane membrane membrane membrane binding to the thymidyl-serine (PS): cells: st.

8, 14 M NaCl and 2.5 µM CaCl 3) were stained with Anuexin V-Atto 4R8. Samples were analyzed on a FAGSGalrbnr eltofluorimeter using CeilQuesE ^M software (Beden, Üllinsoo & Co.).

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Duman: and mouse MSCs (Isit) were grown on round glass slide plates ( ⁴ sep / mfeta). To remove the cell surface GaM, cells were started with SÖÖMM Dodgalactoside (TDG) at 4 ° C for 30 min and then washed extensively with tissue culture medium. The efficiency of Gal-removal was checked by cunion-rimetry. In the TransweW system, hMSCs were placed in the lower vessel, T cells were placed in the upper vessel, and the two cells were selected from nicroporous polyester termerm (0.4 DM). eh T cells (2xi New Hoeehst 33342 were labeled with MSCs for 16 hours in Rx culture). The cells were then fixed for 4 hours at room temperature with 4% formaldehyde and stained for expressing Gal-1d and phosphoridii as follows; , the free protein binding sites on the flap were flushed with FACS buffer, followed by anti-self-monoclonal body mock-up, and in the second step, anti-mouse IgGNL5? and Ánnexln-V-Afexa Fluor 4 & the coverslips were placed on the glass slides with a drop of fixing medium (Fluoromonnt-G).

Samples were examined with a Cari Zeiss (Axioskop: 2Mot) finescence microscope with a magnification of 44x, photographed with an AxiuCam camera and Axíovision 3.1 software, for confophical microscopy we used Oymymns FV1ÖÖÖ laser scanning microscope and oscilloscope software. Adobe Photoshop 71).

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The 2x10 * cells were prepared in 12% SOS pollakrlla.mid gel, and recombinant Gabl was then transferred to a trhrocHeat membrane in TBS containing 0.0.0% Tweea20-af. % gelatin, and then treated with anti-ω-1-Ig (ntAb, 3Cl / A1) and HRP-vd-linked autt-mouse IgG. Goat immunoreactive proteins were visualized using the FCL phts detection system (Amershatn Siosofences).

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The goat RNA and scseramiedj RNA were cloned into the MSe cell wall using the pSttperior RNAi system to obtain Gal-1 silenced and transfected control cells with seRNA.

We used the following primes:

.S'-CjATCCCCACAI'GGAGGCCATC'AACTATTCAAOAGAIACrnGAfGGCCTCCATGITITrrA mGaíee348síL:

5'-AöCTrAAAAAACAT (K3AGG € 'CATCAACTATCTCTfGAÁTí \ GTTGÁTO3CCTCCÁTGrGOG

S'-ÖATCCGCCAaiAACAAATOCACGTGTTrCAAfíAGAACACGTGCAITlGTrCGlGTrrTFA mGalee340scL:

S ^, -A: GCnAAAAACACGAAGAAAKlCAGGTGTTC3rrfGAAAC: AGG'R3CA3''nX3TTCG1OGGG

:: Galeel30sHJ

'5 ^ ATC € CCMACCT' (JrGCCTGCACn'CArrCAAGAGATaAAGT 5C'AGGCACAÖÖITnTr € {A

GslecO0siL:

5'AGCTrAAAAAÁAGCTGTOGCroCAGTlCA'lC'rcTrGAA1GAAGTGCAGGCACAGGTrGGG

Gsiecl20sdJ:

fe-CATCCCCCGTÁeÁCÁTÁeWGCöCTnGAAÖAÖöeÖCAAGTATÖGTÁCGTrFItY & WClA JLZ

S'.AGCTrAAAAACGTACAGATACTOCGCTOTCTreAAAGCÖGAAÖTATOTGTACGGG

The names of primers are the following overlays:

sk short interfering food: rö-RNA (short mterfermg..fe3irpin: mRNA from mouse gaffectin; Gatec :: galectin-d; se scrambled (k-troll with appropriate slRNA); O: upper strand; L: Thread:

more 34 © and i 20: the position of the eagles is the nucleoid, wly 19bp oligonuble otid begins.

The transfected MSGs are selected on the basis of genetically expressed amylotoxicum, and stable cell lineages are identified, their Gal-1 production is characterized by Western biotting, and their T-cell apoptosis is detected in a co-cortical system (Fig. 4). In addition, we determine whether or not grace is maintained

MCS has the features. It considers the expression of MSC markers and dbphenealisation to bone and fat cells.

2. Expression of Gal-l in human and mouse MSC

A. Expression of Gal-l human and mouse MS € -k was amplified in paraffin-embedded .fe ufe-o. human CB .., first column of strain 20 and: mouse (FIG. 2B, second column) mesenchymal stem cells are lysed in SDS sample buffer and 12% - © $ polyacrylamide gel. Purified recombinant Gal-1 was used as a control (Figure 18, third column). Proteins & gels with nitrpellulose; were transferred to a filter and treated with anti-Gal-l Meoluene antibody followed by anti-mouse Ig-HRP. Reactions were visualized on the EGE Boy d.fection system.

The cytofluorimellar assay showed that the hMSCs were squamous to the Gal beta (Figure 2A). Treatment or untreated cells of an effective phthalate analogue Ihtodigalakwzidti &lt; 1 &gt; (I) were stained with anti-Gal-lrsAbv and a second taxi 488-labeled suture mouse IgG and compared to FACS. The effect of TDÖ treatment resulted in a significant decrease in Gal-1 levels on the cell surface (Fig. 2Ά). The relative filament intensity of the samples is described in Materials and Methods

30 · (Fig. 28).

3. Gai-1 secreted by MSC induces apoptosis of T cells

The cytotoxic responsiveness of Gal-1 secreted by MSCs was examined in the T-cell line co-culture system of MSCs and Jutka (Figure 3).

T-cell apoptosis was detected with Annexin V-ÁlexaFluor 4M (Molecuiar Frohes, Invltrogen Girl? 3.5 Lafeta) or · Annexin: V-TRANSMIT: ((ATTO--KC, Mgm-Aidneli). Approximately 3% of T cell death by apeptosis within the first hour of the study was significantly reduced by MSC-induced T-cell apoptosis, indicating the role of the Gal-1 mediator (Figure 3).

Gabbet Net Expressing 1-CellsThis expression occurred during contact with the MSC, as confirmed by the presence of samples with the and-Gal-1 mAb, Fz confirms that Gal-1 is a T- cells were linked to the surface glycoconjugate. The Gal-1 phenotoxic effect of MSC origin was also confirmed by assaying for wild-type, serambled (Gal-15 producer) and Gal-1 siRMS (Gal-1 production) salts MSC cell lines ( * '' and ^'55 ) Inducing effect of T-cell apoptosis, Figure 4 shows that Gabi silenced (siRMS-teastectomy) mMSCT ⁵² and mMS ^Sili cellular eitotoxic typing was greatly reduced with wild-type and seRNA (MSC * '^) compared to fmnSecreted control hox, ΔH-human MSCs (Fig. 3B) and murine MSCs (not shown) also induce active T cell apoptosis, indicating that healthy T cells, like leukemic Jurkat T cells are sensitive to Gal-l-induced apoptosis. In addition, JnrteII T cells also underwent TDG-inhibitory apoptosis in the presence of murine MSCs (Figures 3A and C).

A. In the experiments described above, Gabi produced by MSC has been shown to be a critical factor in tomotokhart T-soap apoptosis.

For the study of the mediated T-cell apoptosis induced by mMSC-derived Gabi mice, IS hMSCs and Jutka cells were co-cultured in such a way that cell-cell interaction could be effected or prevented by a transweil membrane (Figures 5C and D). Hoeehst 33342 stained Jurkat cells (blue or dark gray) were removed and Annenin V-Atto 488 (AsnV, greeu, or light gray, arrowed), or aotböab mosoclonal antibody followed by anti-mouse IgNortheruLights 5S? B and 1.3, red or medium gray), and finally the samples were analyzed with a fluorescent microscope. To determine the T cell apoptosis mice, Aunexln V-Atto 488-labeled Jurkut cells were subjected to FACS analysis in the following samples: Jurkat and: MSC coculture, Transwell membrane-separated Jurkat and MSC cocultures, Jikakat cells are unique to the firasswll insertion. and n 24-hole tissue-printing plates. These results are plotted (Figure 5 £). In the above samples, the percentage of apoptosis was determined by detecting Arsnexin V positive cells.

Based on the results shown in Figure 5, it can be concluded that MSC and T cell defect must be in direct cell-cell interaction in order for Gabi and T cells to undergo apoptosis from the MSC to the T cell membrane. Physical separation of MSCs and T cells results in a lack of Gal-1 'translocation and apoptosis induction. Consistent with this, the conditioned medium from MSC cell culture, as measured by b.USA, did not contain detectable amounts of Gabi (not shown) and accordingly did not induce T cell death (not shown).

Thus, MSU-mediated T-cell apoptosis requires a direct cell-cell rumen effect (Figure 5),

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Mouse MSCs were stably mutated with OI-I siRMS to silence the Gal-1 expression (see Example 1). We have found that the T-cell cytotoxic hamsa of m'MSCs deficient in Gal-1 expression is reduced in the control of transfected with a plasmid containing type 1: d or scrumhlcd RNA.

Your picture for MSCs. Figure 4 shows the theory that a. Qal-Gexpression by down-regulation of RAMSC results in a lower level of apopiotic activity in Jurknt T cells,

5. Examination of the effect of gal ·) deficient Mh € on tumor progression.?? / / · / / Rfcffctes AfÓC / fi ramnro.y & grarafeffíte

In my tour models, Sl 6 murine mammary cell cells or 4T1 murine mammary carcinoma cells were killed by CSzb Stone, lilac Mh / o mice here. Tumor cells are either inoculated with either wild-type or GaM silent MSU. Genetically modified MSCs 1. example, we created it. The optimum ternary MSC ratio, which gives the greatest difference between the control and sample groups, is determined by the inferiority. The macroscopic examination of the Nasal animals includes determination of the size (diameter, weight) of the primary tumor, the location and number of visible nmtasks, and the length of survival (except mice sacrificed for early studies). Metastatic or non-metastatic primary tumors up to the site of transplantation and in parenchymal organs are evaluated by conventional histology, immunohistochemistry (IFI), and also by sitit hybridization (ISII j.). and 2) and targets hypoxia-regulated plaques (flJFl. □ and Gal-1),

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The migration of MSCs into the tumor tissue is monitored by MSC-specific niafragers (CD73, GD105 and CDI33). Migration of MSCs from exogenous male mice into tumor tissue transplanted into female mice: Demostralized by ISfl analysis detecting murine Y chromosome. In our prototype, we successfully labeled the sequence ¥8.ora0-speeifikns Ύ8.Ι0 with digoxigenin and used it to detect male cells in formalin-fixed, paraffin-embedded tissue sections.

Alternatively, when the desired antibodies do not function on paraffin sections, endogenous and transplanted MSCs in the tumor are enriched with the anticoagulant MSC elution medium / magnetic bead method. The total MSC content of the cell population thus generated is assessed by antbMSC antibody (CD7B CD105, Sca-1), eco-micrometry, or fluorescence microscopy, and the proportion of Yplofplontone within the entire MSC population is determined by γ-κ-osteosomal KSfl.

The combination of Y-corraosaurus with FISH and MBC anthem painting gives us the opportunity to discover whether or not the trausplanal MSC. is there a transdifferentiated pathway to tetrasassoidal tissue, such as. cells of the associated vasculature of the siroma or tumor.

In this way, the migration of Gabi defiete, genetically modified MSCs into the tumor, and thus its use in the delivery of antitumor drugs, can be traced.

6. The chemotactic effect of tumor cells on wild-type (vMSQ and Gal silenced MSC-rs. The umehan Ismus of the migration of M8Us into the tumor is not known. Therefore, we use the M migration test of ya et al., Rofe of galectin-1 in ntlgraiion. and mvasonon ol human ghöhfestonta «Source Evening Source .1 Nwowg. 2000-8. IQfe 273] to determine whether <5aM plays a role in this gonadal process.VMSC and Δal-1escadcMítr. sumer ^ pek here in lorteoo urns houraem change MSC k ^ 'vvettvny annual media to hydroxycarbamide «crawling medium and thus block cell division for 24 hours

Subsequently, the cells in the middle of the cell culverte are removed by scraping and a round topcoat covered with tamor cells is placed in this area. The cells were incubated in a medium containing hydroxycarbamld for 48 hours, then the cells were fixed and stained with toluene lamps. The number of cells migrating to the scraped surface and the maximum distance of migration are determined.

81.6 (mepheraoma) and 4T1 (breast carcinoma) cells were inoculated with vMSC and Gal-I silenced

MSG-vci, or none, and tested as described in Example 5.

A methanome 3x1 Irish 816F1Ö mehtnómn cell by inoculation of the dorsal soft veins (SOOl mice with or without co-inoculation of tfe vMSC, Growth with change in solid volume is measured in cup foil and inoculum 20 and inoculum, we calculate it using the formula dAÖxŐ.S, where da is smaller, D is the larger diameter The results are shown in Figure 6Λ.

In the following experiments, 10-week-old female Salhfe mice were sacrificed with an amount of 411 mammary gland cells, with or without ⁵ MSCs, under the skin of the endothelium. The number of tndo20 metaslases is evaluated by counting metasasic wet phos and determining the mass of the cap <68, lower graft). The color gamut is calculated using a two-tailed T test.

Our results undoubtedly showed that MSCs stimulate the growth of non-neo-static 816 melanomas (Fig. 6A) and 4TI smiokaremoma as well as the number of 4T1 metastases {68. figure).

5 By way of comparison, the 816 and 4? tumors are also inoculated with Gal-1 silenced MECs,

Tumor sizes, weights, and number of remnant nodules are determined as described above. Smaller femoral space, tendon mass, or lower metastatic laser, compared to those caused by vMSC, indicate beneficial or therapeutic effects of Gal-I silenced MSC. that the Gala quiet ites does not affect it. MSC-keta to migrate to the tumor, so these cells are a suitable means of transport for pre-tumor encapsulation.

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Jfígiíí & x esyíressrdfetfösrr ASE-fem

K. use of precursor secretory plasmid vectors to synthesize Anginex; for oligonucleotide cloning. Gal-1 expressing and Gal-1-inhibited (silenced, "knocked-dowu") MSC cell lines are transfected with Angfoex. The expression of the generated ' ^V ' Aragon expression of the stable cell colonies was used by Q-PCR and Rls-tag expression / His-tag immunoblot: plating, followed by bv?

7nmnr wdwcccccccccccccccccccccccccccccccccccccccccccccccccccccc

Mice with 1316 and 4T1 Cupid tranxzpfeatálbk yMSC * ' ⁸ and Gal-1 were silenced in the presence and absence of MSC ^W Jeles5 and Examples 5-7' are Serbian asalbal. Inhibition of growth and metastasis is expected.

The present invention provides blackness for the additional treatment of various tumors, namely. and therapy. thereby reducing the risk of side effects.

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Claims (9)

25 Claims 1. A genetically modified mesenchymal stem cell (MSC) that has been modified to be unable to express and / or secrete Galectin-1 (Gal-1), The genetically modified MSC according to claim 1, wherein the expression of Gal-1 is irreversibly inhibited ('knocked out') or 'knocked down'. The genetically modified MSC according to claim 1 or 2, wherein the MSC is capable of expressing and / or expressing a Gte-1 inhibitor, The genetically modified MSC of claim 3 for use in the cell therapy of solid tumors. 35 5. A 3-4. The genetically modified MSC according to any one of claims 1 to 3, wherein the Cal-1 inhibitor is Gal-1 blocking agent selected from Gal-1 derivative, anti-Gal neutralizing agent, its derivatives, and Gal-1 coco-trypsin; oligopepid lectin mimetics, obgopeptide glt-co-mimetics, Gal-1 blocking oligopeptides and polypeptides. She. The genetically modified MSC of claim 5, wherein the Gal-1 blocker is an anginex or Gal-1 biologically inactive former. t. 4-6. The genetically modified MSC for use in the cellular therapy of malignant solid tumors according to any one of claims 1 to 5, wherein the solid malignant tumor is selected from the group consisting of lung cancer, colon and rectal cancer, squamous skin cancer, e.g. ino: na, glioma, thyroid cancer, pancreatic cancer, prostate cancer, kidney cells, breast cancer, myaloma. The genetically modified .MSC for use in the cell therapy of malignant solid tumors according to any one of claims 4-7, including cell therapy as autologous or allogeneic cell therapy, and comprising single or multiple administration of the genetically modified MSC, Use of a genetically modified Gal-1 dexcile MSC according to any one of claims 1 to 3 as a delivery device in a m / v tumor animal model for delivery of a Gal-1 inhibitor to a solid tumor or a tumor microenvironment. 15 The use according to claim 9, wherein the Gal-1 inhibitor is a Gal-1 blocking oligopeptide: or a polypeptide expressed by a genetically modified MSC, and wherein the genetically modified MSC is administered to, delivered to and / or administered to the tumor microenvironment. . migrate and, preferably, integrate into the Unnorse-sided petal. 20 A kit for delivering a Gal-1 blocking oligopeptide or polypeptide to a solid tumor or a tumor supernatant comprising a genetically modified MSC as defined in any one of claims 1-8; and device - introducing a gene encoding a Gal-1 blocking oligopeptide or polypeptide into the genetically modified MSC, thereby expressing the polypeptide in the MSC, and / or - administering or introducing genetically modified MSC into a tumor or in a tumor microenvironment. 12. 1-3, S. The use of a genetically modified Gal-1 deficient MSC according to any one of claims 1 to 6 for the manufacture of a delivery vehicle for delivering a Gal-1 inhibitor to a solid tumor or a microenvironment of a tumor, 13. The use according to claim 12, wherein the Gal-1 inhibitor is a Gal-1 blocking oligopeptide or polypeptide expressed by and genetically modified by the genetically modified MSC. MSC is administered, delivered to and / or administered into the tumor microenvironment. migrate and, preferably, integrate into the needle-associated petal.