WO2013071015A1

WO2013071015A1 - Ex vivo-modified monocytes as local delivery vehicles to treat diseased arteries

Info

Publication number: WO2013071015A1
Application number: PCT/US2012/064305
Authority: WO
Inventors: Ilia Fishbein; Robert J. Levy
Original assignee: The Children's Hospital Of Philadelphia
Priority date: 2011-11-11
Filing date: 2012-11-09
Publication date: 2013-05-16

Abstract

A vehicle for the delivery of at least one therapeutic agent to a site of vascular disease or injury comprises monocytes that comprise the therapeutic agent. Methods for treating a vascular disease or injury comprise isolating a subject's monocytes, modifying the monocytes to comprise at least one therapeutic agent for delivery to a site of vascular disease or injury, and administering the modified monocytes to the subject. Exemplary therapeutic agents include but are not limited to small molecule drugs, proteins, nucleic acids, and vectors encoding a therapeutic gene.

Description

EX WVO-MODIFIED MONOCYTES AS LOCAL DELIVERY VEHICLES

TO TREAT DISEASED ARTERIES

FIELD OF THE INVENTION

This invention relates generally to modified monocytes that are useful as delivery vehicles to provide therapeutic agents to sites of vascular disease or injury. In particular embodiments, the invention relates to methods for treating vascular disease or injury by isolating a subject's monocytes, modifying them to comprise at least one therapeutic agent, and administering the modified monocytes to the subject.

BACKGROUND OF THE INVENTION

Monocyte derived macrophages (MDM) play a central role in the pathogenesis of atherosclerosis, restenosis, and other diseases with a prominent inflammatory component. A vast number of experimental and clinical stud ies over the past twenty years have unequivocally implicated MDM in triggering and maintaining vascular inflammation across a wide scope of diseases and conditions. Furthermore, the extent and activity of atherosclerosis and restenosis have been demonstrated to be linked to

monocyte/macrophage dynamics in both experimental models of vascular disease and in patients. Therefore, it is counter intuitive to consider that MDM could somehow be used for therapeutic purposes.

Millions of angioplasties are performed throughout the world each year. Although there have been improved outcomes over recent years, there is an unmet need for more definite, positive therapeutic results. For example, there is still a 5% instent restenosis incidence in coronary drug eluting stent procedures after one year, affecting hundreds of thousands of patients. Angioplasty for peripheral artery disease (PAD) is only modestly successful with a progressive incidence of 20% annually of restenosis following either balloon angioplasty or stenting for PAD. The deleterious role of macrophages in the natural history of atherosclerotic and restenotic lesions makes intuitively undesirable any extra burdening of experimental subjects or patients with monocytes. This reasoning is probably the main cause for disregarding the possible therapeutic potential of monocytes as delivery vehicles with superb homing capabilities to diseased or injured arteries.

SUMMARY OF THE INVENTION

An embodiment of the present invention provides a vehicle for the delivery of a therapeutic agent to a site of vascular disease or injury (i.e., a therapeutic carrier) comprising monocytes that have been modified to comprise at least one therapeutic agent. The therapeutic agent(s) may comprise, for example, a small molecule drug, a protein, a nucleic acid, a vector encoding a nucleic acid, or a combination thereof. The modified monocytes are capable of providing a therapeutic effect to the site of vascular disease or injury. Another embodiment of the present invention provides a method for treating a vascular disease or injury comprising isolating a subject's monocytes, modifying the monocytes to comprise at least one therapeutic agent for delivery to a site of vascular disease or injury, and administering the modified monocytes to the subject.

BRIEF DESCRIPTION OF THE FIGURES

Figure 1 : Comparison of neointima-to-media area ratio in rats receiving Ad-APN- transduced monocytes, rats receiving Ad-Luc-transduced monocytes, and rats receiving no cell therapy.

Figure 2: Comparison of % stenosis in rats receiving Ad-APN-transduced monocytes, rats receiving Ad-Luc-transduced monocytes, and rats receiving no cell therapy.

DETAILED DESCRIPTION OF THE INVENTION

The applicants have discovered innovative methods that utilize monocytes as therapeutic carriers to target sites of inflammation across a wide array of diseases and conditions. In particular, the modified monocytes of the present invention can provide therapeutic effect to vascular diseases and conditions varying from mild dysfunction to overt injury, including but not limited to atherosclerosis, arteriosclerosis, vasculitis, thromboangiitis obliterans, post-angioplasty/post-stenting restenosis, post-grafting restenosis, and transplantation vasculopathy. The possible therapeutic potential of monocytes as delivery vehicles has previously been disregarded, largely due to the deleterious role of monocytes in the natural history of atherosclerotic and restenotic lesions.

Embodiments of the present invention provide vehicles for the delivery of a therapeutic agent to a site of vascular disease or injury (i.e., a therapeutic carrier), the vehicle comprising monocytes that comprise at least one therapeutic agent (also referred to herein as "modified monocytes"). In an exemplary embodiment, the modified monocytes are autologous monocytes. Preferably, the monocytes that are modified to comprise a therapeutic agent(s) have been isolated from the blood or bone marrow of a subject (i.e., an animal or human) and have not yet differentiated to become

macrophages.

The modified monocytes are preferably targeted to diseased or injured sites in the cardiovascular system, such as angioplastied veins or arteries. For example, the modified monocytes are capable of inhibiting a disease process that affects the cardiovascular system and/or providing a therapeutic effect to angioplastied arteries. The modified monocytes are capable of providing therapeutic effect(s) to a site of, for example, atherosclerosis, arteriosclerosis, vasculitis, thromboangiitis obliterans, post- angioplasty/post-stenting restenosis, post-grafting restenosis, or transplantation vasculopathy. In particular embodiments, the monocytes are modified to exhibit anti- atherosclerotic activity, anti-restenotic activity, or both.

The therapeutic agent(s) carried by the monocytes may comprise any therapeutic agent that is capable of providing a therapeutic effect to a vascular disease or injury. For example, the therapeutic agent may comprise a small molecule drug, a protein, a nucleic acid, a vector encoding a therapeutic gene, or a combination thereof. The delivery vehicle may further comprise any suitable medium or carrier for the modified monocytes. It is not necessary for 100% of the monocytes in the delivery vehicle to comprise the therapeutic agent(s), as long as an effective amount of the monocytes comprise the therapeutic agent(s) to provide a therapeutic effect to a vascular disease or injury.

Preferably, the majority of the monocytes in the delivery vehicle comprise the therapeutic agent(s) .

In particular embodiments, the monocytes are modified to express a secretable transgene product by, for example, transducing the monocytes with a therapeutic gene. Monocyte transduction can provide prolonged or even permanent modification of tissue macrophages derived from the delivered monocytes. The therapeutic agent may comprise a vector that contains a therapeutic gene, such as a viral vector, plasmid DNA, or any other vector known in the art. The list of potential therapeutic transgenes for monocyte expression is extensive and includes, but is not l imited to, adiponectin, AMPK, apoAl , angiotensin converting enzyme, heme oxygenase, 12/15-lipoxygenase, LXR, STAT6, PPARy, and many other candidate genes.

According to some embodiments, monocytes isolated from peripheral blood of subjects, including animals or human patients, are genetically or epigenetically modified ex vivo to bestow temporary or permanent expression of a therapeutic transgene or desired phenotype with therapeutic activity, such as anti-atherosclerotic or anti-restenotic activity. When returned back to the subject's circulation, the modified monocytes consequently become engaged (typically proportional to their representation in the total monocyte population) into sites of ongoing vascular inflammation and express secretable therapeutic transgene product(s), thus altering the inflammatory milieu of atherosclerotic and restenotic lesions. The monocytes themselves, through both autocrine and paracrine actions of the expressed transgene, can become primary targets of the therapeutic intervention .

According to other aspects of the invention, the at least one therapeutic agent comprises any drug capable of providing a therapeutic effect to a vascular disease or injury. One example of such a drug is paclitaxel . The monocytes may be modified by incubating a cell culture of isolated monocytes with at least one drug . In other embodiments, the modified monocytes comprise therapeutic agent(s) that have been loaded into magnetic nanoparticles (MNP) . The therapeutic agent may be included in the MNP as a sustained release preparation. The MNP may contain targeting ligands on their surfaces to enable uptake by the monocytes. The MNP may also be modified with iron oxide to enable magnetically driven loading into the monocytes.

According to some aspects, the monocytes may be modified to inhibit their inflammatory potential. Tissue macrophages, including those in vasculature, exhibit a substantial degree of morphological and functional heterogeneity. It is established that distinct macrophage phenotypes occur as a result of macrophage adaptation to different tasks in local tissue homeostasis. These phenotype changes are mediated by tissue- derived signals that affect macrophage-relevant signaling context in the microenvironment of inflamed tissue. Typically, macrophages are classified either as Ml or M2 depending on the repertoire of expressed molecules and distinct roles played at different stages of tissue inflammation. Classically activated (Ml) macrophages are derived through TNF or combined IFNy and LPS activation of monocytes. They synthesize large quantities of TNF, Ιί β, IL6, IL12, and other inflammatory mediators. The main role of Ml cells is pathogen elimination and tissue destruction. In contrast, alternatively activated (M2) macrophages are obtained by IL4 or IL13 stimulation. These cells synthesize large amounts of antiinflammatory IL10, TGF , and IL1 receptor antagonist, thus opposing the proinflammatory effects of Ml macrophages. Therefore, the main role of M2 cells is curbing inflammatory signaling, allowing resolution of inflammation, and tissue healing. M1/M2 polarization was observed in both experimental and human atherosclerosis. Interestingly, disease progression in mice is associated with a rapid shift in M1/M2 balance toward an accumulation of the Ml pro-inflammatory macrophages. Furthermore, exposure of human monocytes to C-reactive protein (CRP), an inflammatory mediator relevant to human atherosclerosis, primes macrophages toward an Ml differentiation program.

Modified monocytes of the present invention can instigate a differentiation into M2 macrophages preferentially over Ml macrophages. Monocytes may be modified, for example, by reducing the expression of a gene that enhances the monocytes'

inflammatory potential (e.g., by techniques involving RNAi). Alternatively, the monocytes may be incubated with at least one inhibitor. The inhibitor may comprise, for example, a small molecule, a protein, a nucleic acid, or a vector encoding a nucleic acid. The inhibitor may also comprise an antibody that inhibits inflammatory cytokine function, such as a TNF-alpha function blocking antibody or a similar construct for any inflammatory mediator.

Additional embodiments of the present invention provide a cardiovascular device for administering the modified monocytes to a subject. A cardiovascular device for delivering the modified monocytes may comprise, for example, a stent or a catheter, such as a magnetic targeting catheter, as described, for example, in International Application No. PCT/US2011/058029, filed October 27, 2011, the content of which is incorporated by reference herein in its entirety.

Embodiments of the present invention also provide a method for treating a vascular disease or injury comprising isolating a subject's monocytes, modifying the monocytes to comprise at least one therapeutic agent for delivery to a site of vascular disease or injury, and administering the modified monocytes to the subject (e.g., an animal or a human patient). The monocytes may be modified by any of the processes described herein.

In one clinical setting, a patient would have a prepa ration of their own monocytes prepared prior to a procedure and appropriately modified, for example, through transduction with a gene vector, exposure to a pharmaceutical, or magnetic loading with magnetic nanoparticles (MNP). The MNP could contain sustained release pharmaceutics, gene vectors, recombinant proteins, or mixtures of all of these therapeutic entities. In addition, MNP would make the modified monocytes magnetically responsive, and thus adhesion of the modified monocytes could be even further enhanced with magnetic targeting using a magnetic targeting catheter as a delivery system in the presence of a uniform magnetic field.

The step of administering the modified monocytes to the subject preferably comprises administering an amount of the modified monocytes that is effective to provide a therapeutic effect to the site of vascular disease or injury. For example, the modified monocytes may be administered in an amount of about 10⁵ cells/kg to about 10⁷ cells/kg. The modified monocytes can be administered in any suitable medium or carrier. They can be peripherally administered to the subject intravenously or they may be injected locally at the site of an interventional procedure, such as at the site of a coronary stent deployment or balloon angioplasty. The modified monocytes can be administered at any time with respect to the occurrence of the vascular disease or injury. For example, the modified monocytes can be administered at any time after a patient has been diagnosed with a vascular disease, or any time after a patient has undergone an interventional cardiovascular procedure, such as coronary stent deployment or balloon angioplasty. After being administered, monocytes may take several days (e.g., about five to seven days) to arrive at a site of vascular disease or injury. In some embodiments, the modified monocytes are administered about three to seven days after an interventional

cardiovascular procedure.

In particular embodiments, autologous monocytes are used to target sites of vascular disease or injury using a system in which, prior to a cardiovascular intervention, a patient's or animal's own monocytes are isolated either from peripheral blood or from a bone marrow aspirate. The subject then undergoes a cardiovascular interventional procedure, such as stent deployment, balloon angioplasty, or both. The monocytes are then modified to confer either therapeutic capabilities, imaging capabilities, or both. At an appropriate time (e.g., 3-7 days after the invasive procedure), the patient's or animal's monocytes, modified as needed, are injected either locally at the site of intervention, or injected peripherally (e.g., intravenously). It is contemplated that the interventional procedure can be performed before or after any of the steps of isolating the monocytes, modifying the monocytes, and re-administering the modified monocytes.

Additional embodiments of the present invention provide methods for modifying a subject's monocytes to comprise at least one therapeutic agent for delivery to a site of vascular disease or injury according to any of the processes described herein.

The following examples are provided to describe embodiments of the invention in greater detail and are intended to illustrate, not limit, the invention.

EXAMPLES

Example 1. MDM homing to injured arteries.

Fogarty balloon catheter denudation injury was modeled in carotid arteries of 2 male Sprague-Dawley rats (400-420 g). Four days after the surgery, 8 ml of blood was harvested at sacrifice of an additional naive rat. Blood was mixed with 4 ml PBS and was spun on Ficoll gradient (Accuspin; Sigma-Aldrich). Plasma was discarded and the buffy coat layer was collected, diluted in PBS and washed by centrifugation in PBS, The pellet was then exposed to 3 ml ACK buffer on ice for 5 minutes, diluted to 30 ml with PBS, divided into 2 tubes and spun again for 12 minutes at 1000 g (large centrifuge). The pellets were suspended then into 5 ml volumes of 2 μΜ CM-Dil to label monocytes, and incubated for 5 minutes at 37°C and for additional 15 minutes at 4°C.

The cells were then washed twice in PBS and were suspended in 2 ml of

20%FBS/1%PS RPMI-1600. Cells were counted in a hemocytometer (3.5xl0⁵/ml). Within 30 minutes of cell isolation, 1 ml aliquots of labeled mononuclear cells were injected IV to the rats that underwent Fogarty procedure 4 days earlier.

The rats were euthanized 8 days after the surgery and 4 days after the cell transplantation. The harvested arteries were fixed in formalin for 24 hours, equilibrated in sucrose, OCT embedded, cut (10 pm), counterstained with DAPI, and examined by fluorescence microscopy. Labeled cells were ubiquitously present in the nascent neointima, demonstrating the ability of monocytes to target a site of vascular injury after they have been administered to a subject.

Example 2. MDM homing and persistent transgene activity to injured arteries.

Fogarty balloon catheter denudation injury was modeled in carotid arteries of 4 male Sprague-Dawley rats (400-450 g). Three days later 12 ml of peripheral blood was harvested in heparinized syringe at the sacrifice of additional rat. Blood was layered on Ficoll-Paque Premium (Fisher Scientific) and centrifuged at 700 g for 30 minutes. Buffy coat layer was collected, diluted with PBS to 15 ml and layered on Percoll (Fisher Scientific), followed by the repeated gradient centrifugation (700 g, 30 minutes). The buffy coat layer containing white blood cells was collected and washed twice in 1%

BSA/PBS. The cells were suspended in 22.5 ml of 10% FBS/ RPMI1640, and 100 μΙ of AdLuc (4.99xl0¹²/ml) were added to the cells followed by centrifugation of cells and virus for 80 minutes at 2000 g (according to Mayne et al; J. Immunol Methods; 2003, 278 : 45- 56). The cells were re-suspended in 10% FBS/ RPMI1640 and placed overnight into T-150 flask. Next day the cells were washed to remove non-attached cells. The attached cells were trypsinized, collected by centrifugation re-suspended in 4 ml of 10% FBS/ RPMI1640 and counted. Within 30 minutes of cell preparation 1-ml aliquots were injected IV to the animals that underwent Fogarty procedure 4 days prior to cell injections.

Twenty-four hours after cell injection, the animals were imaged for 5 minutes after the local application of 500pg luciferin formulated as Pluronic/luciferin mixture (4: 1) to the balloon-injured artery. The animals were recovered and then reimaged on day 3 after cell therapy. In all animals, the signal emitting from the injured artery was found at 1 day time point. A signal at day 3 was attenuated compared to the 1-day assessment. Thus, these results demonstrate the concept of using MDM as therapeutic carriers that can be self-targeted to angioplasty-injured sites. This approach would be particularly useful after interventional procedures such as coronary stent deployment, or balloon angioplasty, commonly performed for peripheral arterial disease.

Example 3,

Fogarty denudation injury was modeled in left carotid arteries of 16 male Sprague- Dawley rats. Two days after surgery 12 out of 16 rats received IV injections of 7.5xl0⁵ monocyte-enriched cells transduced with either Ad-APN (n=6) or Ad-Luc (n=6) as described below. Remaining 4 rats were not treated beyond vascular denudation injury.

Twenty ml of blood was harvested from healthy littermates. Blood was diluted 1 : 1 with PBS and buffy coats (peripheral blood mononuclear cell fraction) were obtained by Ficoll centrifugation. The cells were washed twice in PBS and exposed to ACK buffer to lyse contaminating erythrocytes, re-suspended in 1% BSA/1 mM EDTA/PBS and washed twice by centrifugation at 200G to eliminate platelets. The resulting pellet (9.2xl0⁷ cells) was re-suspended in 1.5 ml of 1% BSA/1 mM EDTA/PBS on ice. Twenty g of each of the following mouse anti-rat antibodies [anti-CD5 (OX19), anti-CD6 (0X52), anti-CD8a (OX8) and anti-CD45RA (0X33)] were added to cell suspension. This anti-lymphocyte antibody cocktail binds T-, and B-lymphocyte as well as NK cells, whi le sparing monocytes. After 1 hour incubation at 4°C with mild shaking the cells were washed twice by centrifugation and re-suspended in 800 μΙ of chilled 1% BSA/1 mM EDTA/PBS buffer. One hundred and forty μΙ of anti-mouse IgG antibody conjugated to magnetic beads (Miltenyi Biotech) were added to cell suspension and incubated at 4°C for 15 minutes followed by centrifugation, re-suspension in 1 ml of 1% BSA/1 mM EDTA/PBS buffer, and magnetic separation using a LS column (Miltenyi Biotech). The cell fraction eluted from the column (9xl0⁶ cells total) was concentrated by centrifugation, re-suspended in complete RPMI-1640 medium, and divided into 2 aliquots of 5 ml each. These aliquots were injected into sterile Teflon bags and were transduced with 50 μΙ of either Ad-Luc or Ad-APN for 12 hours at 37°C. Finally the cells were washed twice by centrifugation to remove free virus, resuspended in complete RPMI-1640 medium at a concentration of 7.5xl0⁵/inl, passed through 70 pm nylon mesh to eliminate clumps of cells, and injected to anesthetized rats via tail vein.

The rats were euthanized 14 days after surgery. The injured carotid arteries were harvested, formalin-fixed, embedded in OCT medium, cryosectioned, and stained by modified Verhoef-VanGieson method.

Computerized morphometry demonstrated reduction of restenosis indices (% of stenosis and neointima-to-media area ratio) in rats receiving Ad-APN-transduced monocytes in comparison with the animals treated with Ad-Luc-transduced monocytes. The computerized morphometry also demonstrated reduction of restenosis indices in rats receiving Ad-APN-transduced monocytes in comparison with the rats that underwent Fogarty balloon injury without cell therapy (Figures 1 and 2).

Although the present invention has been described in connection with specific embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications and variations of the described compositions and methods of the invention will be apparent to those of ordinary skill in the art and are intended to be within the scope of the appended claims.

Claims

CLAIMS What is claimed :

1. A vehicle for the delivery of at least one therapeutic agent to a site of vascular disease or injury comprising monocytes that comprise the at least one

therapeutic agent.

2. The delivery vehicle of claim 1, wherein the monocytes are autologous monocytes.

3. The delivery vehicle of claim 1, wherein the monocytes exhibit anti- atherosclerotic activity, anti-restenotic activity, or both.

4. The delivery vehicle of claim 1, wherein the monocytes preferentially differentiate into a macrophage having an M2 phenotype over an Ml phenotype.

5. The delivery vehicle of claim 1, wherein the monocytes express a secretable transgene product.

6. The delivery vehicle of claim 1, wherein the monocytes are capable of providing a therapeutic effect to the site of vascular disease or injury.

7. The delivery vehicle of claim 1, wherein the monocytes are capable of inhibiting a disease process that affects the cardiovascular system.

8. The delivery vehicle of claim 1, wherein the monocytes are capable of providing a therapeutic effect to angioplastied arteries.

9. The delivery vehicle of claim 1, wherein the monocytes are capable of providing a therapeutic effect to a site of atherosclerosis, arteriosclerosis, vasculitis, thromboangiitis obliterans, post-angioplasty/post-stenting restenosis, post-grafting restenosis, or transplantation vasculopathy.

10. The delivery vehicle of claim 1, wherein the at least one therapeutic agent is selected from the group consisting of a small molecule drug , a protein, a nucleic acid, a vector encoding a therapeutic gene, and a combination thereof.

11. The delivery vehicle of claim 1, wherein the at least one therapeutic agent comprises a vector containing a therapeutic gene.

12. The delivery vehicle of claim 11, wherein the therapeutic gene is selected from the group consisting of adiponectin, AMPK, apoAl, angiotensin converting enzyme, heme oxygenase, 12/15-lipoxygenase, LXR, STAT6, and PPARy.

13. The delivery vehicle of claim 11, wherein the vector comprises a viral vector or plasmid DNA.

14. The delivery vehicle of claim 1, wherein the at least one therapeutic agent comprises a drug.

15. The delivery vehicle of claim 14, wherein the drug comprises paclitaxel.

16. The delivery vehicle of claim 1, wherein the at least one therapeutic agent is loaded into magnetic nanoparticles (MNP).

17. The delivery vehicle of claim 16, wherein the at least one therapeutic agent is included in the MNP as a sustained release preparation.

18. The delivery vehicle of claim 16 , wherein the MNP contain targeting ligands on their surfaces to enable uptake by the monocytes

19. The delivery vehicle of claim 16 , wherein the MNP are modified with iron oxide to enable magnetically driven loading into cells.

20. The delivery vehicle of claim 1, wherein the monocytes have been modified to inhibit their inflammatory potential.

21. A cardiovascular device for administering the delivery vehicle of claim 1 to a subject.

22. A cardiovascular device comprising the delivery vehicle of claim 1.

23. The cardiovascular device of claim 21 comprising a catheter or a stent,

24. The cardiovascular device of claim 21 comprising a magnetic targeting catheter.

25. A method for treating a vascular disease or injury comprising : isolating a subject's monocytes; modifying the monocytes to comprise at least one therapeutic agent for delivery to a site of vascular disease or injury; and administering the modified monocytes to the subject.

26. The method of claim 25, wherein the modified monocytes are capable of providing a therapeutic effect to the site of vascular disease or injury.

27. The method of claim 25, wherein the modified monocytes are capable of inhibiting a disease process that affects the cardiovascular system.

28. The method of claim 25, wherein the modified monocytes are capable of providing a therapeutic effect to angioplastied arteries.

29. The method of claim 25, wherein the modified monocytes are capable of providing a therapeutic effect to a site of atherosclerosis, arteriosclerosis, vasculitis, thromboangiitis obliterans, post-angioplasty/post-stenting restenosis, post-grafting restenosis, or transplantation vasculopathy.

30. The method of claim 25, wherein the modifying step comprises modifying the monocytes to exhibit anti-atherosclerotic activity, anti-restenotic activity, or both.

31. The method of claim 25, wherein the modifying step comprises modifying the monocytes to preferentially differentiate into a macrophage having an M2 phenotype over an Ml phenotype.

32. The method of claim 25, wherein the at least one therapeutic agent is selected from the group consisting of a small molecule drug, a protein, a nucleic acid, a vector encoding a nucleic acid, and a combination thereof.

33. The method of claim 25, wherein the modifying step comprises transducing the monocytes with a therapeutic gene.

34. The method of claim 25, wherein the modifying step comprises modifying the monocytes to express a secretable transgene product.

35. The method of claim 25, wherein the at least one therapeutic agent comprises a vector containing a therapeutic gene.

36. The method of claim 35, wherein the therapeutic gene is selected from the group consisting of adiponectin, AMPK, apoAl, angiotensin converting enzyme, heme oxygenase, 12/15-lipoxygenase, LXR, STAT6, and PPARy.

37. The method of claim 35, wherein the vector comprises a viral vector or plasmid DNA.

38. The method of claim 25, wherein the at least one therapeutic agent comprises a drug.

39. The method of claim 38, wherein the drug comprises paclitaxel.

40. The method of claim 38 comprising incubating a cell culture of the isolated monocytes with the at least one drug.

41. The method of claim 25, wherein the at least one therapeutic agent is loaded into magnetic nanoparticles (MNP).

42. The method of claim 41 , wherein the at least one therapeutic agent is included in the MNP as a sustained release preparation.

43. The method of claim 41, wherein the MNP contain targeting ligands on their surfaces to enable uptake by the monocytes.

44. The method of claim 41, wherein the MNP are modified with iron oxide to enable magnetically driven loading into cells.

45. The method of claim 25, wherein the modifying step comprises modifying the monocytes to inhibit their inflammatory potential.

46. The method of claim 45, comprising reducing the expression of a gene that enhances the monocytes' inflammatory potential (e.g., RNAi).

47. The method of claim 45 comprising incubating the monocytes with at least one inhibitor.

48. The method of claim 47, wherein the at least one inhibitor comprises a small molecule, a protein, a nucleic acid, or a vector encoding a nucleic acid.

49. The method of claim 47, wherein the at least one inhibitor comprises an antibody that inhibits inflammatory cytokine function, such as a TNF-alpha function blocking antibody.

50. The method of claim 25, wherein the administering step comprises administering an effective amount of the modified monocytes to provide a therapeutic effect to the site of vascular disease or injury.

51. The method of claim 25, wherein the administering step comprises administering the modified monocytes in an amount of about 10⁵ cells/kg to about 10⁷ cells/kg.

52. The method of claim 25, wherein the administering step comprises injecting the modified monocytes into the subject intravenously.

53. The method of claim 25, wherein the administering step comprises injecting the modified monocytes locally at the site of an interventional procedure.

54. The method of claim 53, wherein the interventional procedure is a coronary stent deployment or a balloon angioplasty.

55. The method of claim 25, wherein the administering step comprises administering the modified monocytes about three to seven days after an interventional procedure.