WO2007123851A2 - Procédé et appareil pour modifier la conduction électrique dans le coeur au moyen de cellules souches sanguines à activité profibrotique - Google Patents

Procédé et appareil pour modifier la conduction électrique dans le coeur au moyen de cellules souches sanguines à activité profibrotique Download PDF

Info

Publication number
WO2007123851A2
WO2007123851A2 PCT/US2007/009168 US2007009168W WO2007123851A2 WO 2007123851 A2 WO2007123851 A2 WO 2007123851A2 US 2007009168 W US2007009168 W US 2007009168W WO 2007123851 A2 WO2007123851 A2 WO 2007123851A2
Authority
WO
WIPO (PCT)
Prior art keywords
pfpcs
source
heart
differentiation
cells
Prior art date
Application number
PCT/US2007/009168
Other languages
English (en)
Other versions
WO2007123851A3 (fr
Inventor
James William Broderick
Frank A. Ahmann
Original Assignee
Symphony Medical, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Symphony Medical, Inc. filed Critical Symphony Medical, Inc.
Publication of WO2007123851A2 publication Critical patent/WO2007123851A2/fr
Publication of WO2007123851A3 publication Critical patent/WO2007123851A3/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/34—Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M25/00—Catheters; Hollow probes
    • A61M25/0067—Catheters; Hollow probes characterised by the distal end, e.g. tips
    • A61M25/0082—Catheter tip comprising a tool
    • A61M25/0084—Catheter tip comprising a tool being one or more injection needles
    • A61M2025/0089—Single injection needle protruding axially, i.e. along the longitudinal axis of the catheter, from the distal tip
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M25/00—Catheters; Hollow probes
    • A61M25/10—Balloon catheters
    • A61M2025/1043—Balloon catheters with special features or adapted for special applications
    • A61M2025/105—Balloon catheters with special features or adapted for special applications having a balloon suitable for drug delivery, e.g. by using holes for delivery, drug coating or membranes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M25/00—Catheters; Hollow probes
    • A61M25/10—Balloon catheters
    • A61M2025/1043—Balloon catheters with special features or adapted for special applications
    • A61M2025/1086—Balloon catheters with special features or adapted for special applications having a special balloon surface topography, e.g. pores, protuberances, spikes or grooves

Definitions

  • the present invention relates to treatment of medical conditions of the heart, and more particularly to treatment of heart arrhythmias and other cardiac conditions by modification of electrical conduction in the heart with bone marrow derivative.
  • Atrial fibrillation is the most frequently occurring sustained cardiac arrhythmia, particularly among the elderly and patients with organic heart disease; and is one of the fastest growing segments of cardiovascular disease in the U.S.
  • Conventional therapies center around ablation (destruction) of the aberrant conduction pathways, accompanied by the implantation of implantable cardiac devices such as defibrillators and pacemakers. Implantable cardiac devices are reasonably effective but have high costs and often undesirable side effects.
  • the apparatus and methods of the present invention advantageously provide for the production and introduction of a therapeutically effective amount of profibrotic blood progenitor cells into regions of the heart for treatment of aberrant conduction pathways.
  • FIG. 1A is a schematic diagram showing one technique for producing and injecting autologous PFPCs into the heart of a patient for conduction modification.
  • FIG. 1B is a schematic diagram showing another technique for producing and injecting autologous PFPCs into the heart of a patient for conduction modification.
  • FIG. 2 is a schematic diagram of the heart showing one approach for introducing PFPCs into the heart for conduction modification.
  • FIG. 3 is a schematic diagram of the heart showing another approach for introducing PFPCs into the heart for conduction modification.
  • FIG. 4 is a schematic diagram of the heart showing another approach for introducing PFPCs into the heart for conduction modification.
  • FIG. 5 is a schematic diagram of the heart showing another approach for introducing PFPCs into the heart for conduction modification.
  • FIG. 6 is a schematic diagram showing one type of injection catheter.
  • FIG. 7 is a schematic diagram showing another type of injection catheter.
  • FIG. 8 is a schematic diagram showing one technique for mixing components of an agent.
  • FIG. 9 is a schematic diagram showing another technique for mixing components of an agent.
  • FIG. 10 is a schematic diagram showing a reentrant conduction pathway.
  • FIG. 11 is a schematic diagram showing injection of agent into the reentrant conduction pathway of FIG. 10.
  • FIG. 12 is a schematic diagram showing correction of the reentrant conduction pathway of FIG. 10.
  • FIG. 13 is a perspective diagram showing an injection system that provides a circumferential pattern for delivery of material.
  • FIG. 14 is a schematic diagram showing a variation of the injection system of FIG. 13.
  • FIG. 15 is a schematic diagram showing a variation of the injection system of FIG. 13.
  • a system forms a conduction block in a regions of cardiac tissue at a location associated with a cardiac arrhythmia by delivering a material that is non-ablative into the region.
  • the material may be living cells that interfere with the normal gap junctions between cardiomyocytes to conduct, such as, for example, myoblasts, stem cells, and fibroblasts.
  • the material may be a non-living agent, such as a polymer agent, e.g. fibrin glue agent or collagen agent.
  • the material may be a combination of living and non-living material that enhances the cellular conduction block.
  • a contact member delivers the material over a patterned region of tissue, such as arcuate, linear, or circumferential patterns.
  • the contact member may include an expandable member or balloon.
  • a guidewire may be used for delivery.
  • Cells used may be autologous, prepared for injection with a kit. Conduction blocks are thus formed without substantially ablating cardiac tissue in the region.
  • Cardiac tissue may have an infarct zone that is related to a reentrant conduction pathway associated with cardiac arrhythmia.
  • the distal end portion of a catheter may be delivered to a location associated with the reentrant pathway, which may be done by, for example, using a mapping electrode provided at the distal tip and via an external mapping or monitoring system coupled to proximal end portion of the catheter outside of the body.
  • This highly localized injection of material across the reentrant circuit blocks the circuit, so that its arrhythmic effects diminish or are entirely remedied with return to sinus rhythm.
  • each type of cardiac arrhythmia may present unique circumstances, both anatomically and functionally, that may in some circumstances benefit from specially adapted delivery devices and techniques.
  • a non-ablative circumferential conduction block may be appropriate to treat atrial arrhythmia, such as forming a circumferential conduction block in a circumferential region of tissue at a location where the pulmonary vein extends from the atrium.
  • the location may be generally at a funneling region or ostium between the atrium and respective pulmonary vein, but may be located up along the pulmonary vein wall itself to the extent cardiac tissue is located there, and may also be considered to include a region of tissue along the back wall of the atrium and closely surrounding the pulmonary vein ostium.
  • All of these regions together may be included in a treatment and may be considered at a "location where a pulmonary vein extends from an atrium," or such treatment may be more localized to only one such place, in which case it is still considered a "location where a pulmonary vein extends from an atrium.”
  • transecting a portion of a region of tissue may be sufficient to block an arrhythmic conduction path therethrough, such as across "fingers” of cardiac tissue that have been observed to extend up from atria and into the base of pulmonary veins. Injectates may be overlapped and partially rotated one or more times for better circumferential coverage and overlapping.
  • a complete or substantially complete circumferential conduction block at a pulmonary vein ostial location may be highly beneficial and optimal.
  • atrial fibrillation may be treated without the need for mapping so extensively to identify which specific vessel houses a focal origin of such arrhythmia.
  • Cardiac conduction disturbances may also be treated by delivery of material into the atrio-ventricular node or the sino-atrial node.
  • Progenitor Cells are advantageous for the non-ablative treatment of conduction disturbances in the heart.
  • Concentrated autologous PFPCs have the ability to modify the electrical pathways of the heart in a variety of different ways, including by spreading conduction as well as by conduction block as scar tissue is formed in response to secretion of collagen, pro-fibrotic growth factors, and cytokines.
  • electromechanical coupling of fibroblasts can be induced, PFPCs may also be used to create new pathways to normalize the conduction of the heart from abnormal conduction pathways.
  • Other approaches may be employed, including the production of gap junction proteins by means of PFPCs for the purpose of normalizing the conduction pathway via electromechanical coupling with the existing cardiac myoctyes.
  • PFPCs and/or other bone marrow lineage cells circulate in the peripheral blood and naturally traffic to wound sites where they participate in the wound healing process
  • circulating PFPCs may be harvested and expanded ex vivo for therapeutic re-administration to the heart in an agent that may or may not contain other therapeutically effective components, including components that promote cell differentiation.
  • PFPCs may be introduced in accordance with various non-ablative methods, apparatus (including systems) and compositions of matter for controlling abnormal heart conduction, such as those described in the '740 published application.
  • PFPCs may be directly introduced into the myocardium such as by injection during open chest procedures, or epicardially or endocardially through a catheter inserted percutaneously or transvenously.
  • PFPCs may be produced in a variety of different ways from a patient's whole blood. Illustrative techniques include the in vitro culturing of PFPCs that have been isolated from the blood, and the differentiation into PFPCs of bone marrow lineage cells that are isolated from the blood. PFPCs may be used immediately or cyro-preserved for later use. PFPCs may be introduced directly after isolation, or after isolation and in vitro culture with or without the introduction of exogenous genes, and with or without expression in culture. PFPCs may be combined with other materials, living or non-living, during injection, such as in the injector or in the tissue at the injection site.
  • PFPCs When introduced into the heart, PFPCs are a strong producer of collagen and is a particularly prolific producer of cytokines, which are important in the healing process. PFPCs also facilitate collagenesis. PFPCs and its secretions are effective in causing modifications to electrical conduction in the heart. [0030] Identification and Function of PFPCs
  • profibrotic cells such as myofibroblasts and fibroblasts are known to be of hematopietic origin and/or circulate in the peripheral blood. In response to inflammatory events surrounding tissue injury, these precursors are recruited to the site of injury where they mature to profibrotic cells such as myofibroblasts and fibroblasts. There, they secrete fibrogenic growth factors and facilitate collagenesis. The exact taxonomy of many of these profibrotic cells and their precursors is still being developed. For our purposes, the taxonomic name given these cells and the precise cell type or types from which they actually come is not important.
  • DMF myofibroblasts and/or fibroblasts, as well as their precursors that are major drivers in fibrosis, derive from adult stem cells, bone marrow cells, and/or other peripheral blood cells, and can be found in circulation in the peripheral blood. Precisely how DMFs differentiate from adult stem cells, bone marrow or bone marrow lineage cells, or other peripheral blood cells is not important for our purposes, and the ultimate taxonomy of these cells as myofibroblasts, fibrocytes or otherwise is not important for our purposes.”
  • Fibrocytes have been described in many publications, including
  • PFPCs have some properties in common with fibroblasts, which are involved In the classic descriptions of wound repair.
  • wound repair begins when the host response is initiated immediately upon the traumatic disruption of tissue.
  • platelets aggregate at the wound site and secrete thrombin, which catalyzes blood clot formation to prevent blood loss, as well as chemoattractants to attract other cells into the damages area.
  • thrombin catalyzes blood clot formation to prevent blood loss, as well as chemoattractants to attract other cells into the damages area.
  • thrombin catalyzes blood clot formation to prevent blood loss, as well as chemoattractants to attract other cells into the damages area.
  • thrombin catalyzes blood clot formation to prevent blood loss, as well as chemoattractants to attract other cells into the damages area.
  • chemoattractants to attract other cells into the damages area.
  • blood-borne neutrophils and monocytes migrate into the wound site and
  • Monocytes mature into macrophages in this environment, which secrete additional chemoattractants such as inflammatory cytokines (tumor necrosis factor, interleukins, transforming growth factory-beta, and so forth), and growth factors such as epidermal growth factor, fibroblast growth factor, and platelet-derived growth factor, which promote immunity, fibroblast activation and replication, and angiogenesis.
  • chemoattractants such as inflammatory cytokines (tumor necrosis factor, interleukins, transforming growth factory-beta, and so forth), and growth factors such as epidermal growth factor, fibroblast growth factor, and platelet-derived growth factor, which promote immunity, fibroblast activation and replication, and angiogenesis.
  • cytokines tumor necrosis factor, interleukins, transforming growth factory-beta, and so forth
  • growth factors such as epidermal growth factor, fibroblast growth factor, and platelet-derived growth factor, which promote immunity, fibroblast activation and replication,
  • fibroblasts are cells of mesenchymal origin while PFPCs and/or their lineage cells circulate in the peripheral blood and express cell surface markers indicative of a hematopoetic origin. While the chemokine expression profile of PFPCs more closely resembles that of activated fibroblasts rather than monocytes or macrophages, they are a significantly more potent source of cytokines, growth factors and matrix components.
  • PFPCs hematogenously enter tissue, where they produces matrix proteins such as vimentin, collagens I and III, and participate in the remodeling response by secreting matrix metalloproteinases.
  • PFPCs are also a rich source of inflammatory cytokines, growth factors, and chemokines that provide important intercellular signals within the context of the local tissue environment.
  • PFPCs express the immunological markers typical of an antigen-presenting cell, and is fully functional for the presentation of antigen to native T cells. Some PFPCs are believed to further differentiate, and may represent a systemic source of the contractile myofibroblast that appears in many fibrotic lesions.
  • PFPCs are found in peripheral blood and quickly appear at wound sites, generally within three days or so. The PFPCs may also play a critical role when fibroblasts cannot contribute significantly to the wound repair process. PFPCs have a variety of distinct functions which make them particularly effective in the wound healing process. PFPCs are capable of presenting Ag to T cells, secreting angiogenic factor to promote new blood vessel growth, and expressing myofibroblast proteins important in contracting collagen for wound closure. PFPCs are believed to be present early in the inflammatory response to present Ag and is also retained at the same site for the resolution of an inflammatory lesion.
  • PFPCs produce a variety of cytokines that can serve to coordinate the successive inflammatory and reparative responses, and is an especially abundant source of these cytokines, as well as of growth factors and matrix components.
  • PFPCs express high levels of the fibrogenic growth factors, PDGF-A and TGF- ⁇ 1.
  • PFPCs also express detectable mRNAs for the pro-inflammatory cytokines, IL-1 ⁇ , but relatively little message for the mediators MIF or TNFa. Collagen production is tightly regulated in the context of the inflammation and wound healing responses.
  • IL- 1 ⁇ for example, induces secretion of the inflammatory chemokines MIP-Ia, MIP-1 ⁇ , MCP-1, IL-8 and GrOa, the hematopoietic growth factors IL-6 and M-CSF, and the fibrogenic cytokines TGF- ⁇ 1 and TNFa.
  • PFPCs migrate early into wounds, where they exhibits a regulated production of both matrix proteins and growth regulating cytokines.
  • PFPCs can express ⁇ -smooth muscle actin, and readily contracts collagen-gels in vitro.
  • PFPCs produce angiogenic factors and may contribute importantly to new blood vessel formation.
  • Expression of matrix metalloproteinase-9 (MMP-9) may further facilitate the angiogenesis process.
  • VEGF vascular endothelial growth factor
  • PDGF-A platelet-derived growth factor A
  • M-CSF macrophage- colony stimulating factor
  • HGF hepatocyte growth factor
  • GM-CSF granulocyte- macrophage-colony stimulating factor
  • CNTGF connective tissue growth factor
  • Other cytokines that have been detected include interleukin-1 ⁇ (IL-1 ⁇ ) and interieukin-8 (IL-8).
  • PFPCs have antigen- presentation properties and may re-stimulate tetanus toxoid-specific T cell responses. Antigen-primed PFPCs may migrate to regional lymph nodes, and to prime native T cells in a Class ll-specific fashion.
  • PFPCs express several chemokine receptors, such as CCR3, CCR5, CCR7, and CXCR4.
  • Peripheral blood PFPCs express the CD34 cell surface antigen.
  • Additional cell surface markers that are indicative of the hematologic origin of PFPCs include CD11b, CD45, HLA-DR, CD71, CD80, and CD86. Additional markers include Type I pro-collagen, the pan-leukocyte antigen CD45, and HLA-DR. Other markers of connective tissue matrix production, such as vimentin, and prolyl 4- hydroxylase, may also be employed.
  • SAP serum amyloid P
  • CRP major acute-phase protein
  • SAP and CRP are secreted by the liver and circulate in the plasma as pe ⁇ tamers.
  • Purified SAP is available commercially from various sources, including the Sigma-Aldrich Company of St. Louis, Missouri, and from EMD Biosciences Inc. (Calbiochem) of San Diego, California. The ability of SAP to inhibit differentiation suggests that its presence in peripheral tissues would be a means of regulating monocyte to fibrocyte differentiation, for example.
  • SAP small changes in the concentration of SAP have a large effect on fibrocyte differentiation.
  • SAP is likely to be either absent or rendered incapable of inhibiting monocyte-fibrocyte differentiation.
  • One possible mechanism at the resolution of an inflammatory response may be that local effective levels of SAP may be reduced due to the binding to DNA and chromatin from dead or damaged cells, thereby enhancing the local differentiation of fibrocytes and improving wound healing.
  • the underlying ECM protein composition is altered, which is likely to affect SAP localization.
  • elevating SAP levels may deter remodeling.
  • Pilling et al. describe a particularly rapid technique for culturing CD14+ peripheral blood monocytes in the absence of serum and plasma to produce fibrocytes within about 72 hours.
  • Peripheral blood monocytes are isolated from a patient's blood using, for example, the Buffy Coat method.
  • SAP is depleted from the plasma using agarose beads (Bio-Gel A), or protein G beads coated with anti-SAP Abs.
  • Agarose depletion of plasma is performed by diluting the plasma to 20% in 10 mM Tris, pH 8, 150 mM NaCI, and 5 mM CaCfc buffer, and mixing with an equal volume of Bio-Gel A-1.5m agarose beads (Bio-Rad, Hercules, CA) for 2 hours at 4oC. Beads are then removed by centrifugation, and the process is repeated twice. The peripheral blood monocytes at 2.5 x 10 5 /ml are then incubated in serum-free medium in the presence of dilutions of plasma at, for example, 0.3% or 0.6%. Ab depletion of plasma is performed using protein G-coated beads (Roche, Indianapolis, IN).
  • a total of 500 ul of protein G beads is coated with a mixture of 20 ⁇ g monoclonal (Chemicon) and 300 ⁇ l of 1/50 rabbit polyclonal antisera (BioGenesis) anti-SAP Abs, in accordance with the manufacturer's instructions.
  • a total of 500 ul of plasma, pretreated at 56°C for 30 min, is diluted to 20% in 20 mM sodium phosphate buffer and precleared with 100 ⁇ l of unlabeled protein G beads for 60 min at 4oC. Beads are then removed by centrifugation. SAP is then depleted from plasma by overnight incubation at 4oC with 100 ⁇ l anti-SAP beads. Beads are removed by centrifugation before cell culture. After depletion, plasma is diluted to 0.25% and cultured with the peripheral blood monocytes at 2.5 x 10 5 /ml.
  • blood-borne mesenchymal cells may be isolated by separation based on the presence or absence of specific cell surface markers.
  • These techniques may include flow cytometry using a fluorescence activated cell sorter or biotin-avidin or biotin-streptavidin separations using biotin-conjugated to marker-specific polyclonal or monoclonal antibodies and avidin or streptavidin bound to a solid support such as affinity column matrix or plastic surfaces, magnetic separations using antibody- coated magnetic beads, destructive separations such as antibody plus complement or antibody coupled to cytotoxins or radioactive isotopes for the removal of undesirable cell populations.
  • mesenchymal cells alternatively may be isolated by procedures involving repetitive density gradient centrifugation, lectin chromatography, affinity chromatography involving positive selection and negative selection, or a combination thereof.
  • Positive selection methods may utilize affinity chromatography with antibodies directed to mesenchymal cell-specific surface markers. For example, most mononuclear cells may be depleted first from the blood after density gradient centrifugation and plastic adhesion, then an antibody to vimentin antigen can be used to positively select for mesenchymal cells.
  • Negative selection includes modifications of the protocol disclosed herein, infra. In essence, a mesenchymal cell preparation may be reacted with one or more antibodies directed at cell surface antigens not expressed by mesenchymal cells for their removal.
  • Antibodies to any T cell, B cell, monocyte, natural killer (NK) cell, dendritic cell and granulocyte markers may be used.
  • examples of such antibodies include anti-CD3, anti-CD4, anti-CD5, anti-CD8, anti-.alpha..beta. and anti-.gamma.. delta. T cell receptor specific for T cells; anti-CD12, anti-CD19 and anti-CD20 specific for B cells; anti-CD14 specific for monocytes; and anti-CD16, and anti-CD56 specific for NK cells.
  • These antibodies may be applied in any combination repeatedly or in a sequential manner for the enrichment of mesenchymal cells.
  • the cells may be removed by adsorption to a solid surface coated with an anti-mouse antibody column, as the majority of monoclonal antibodies directed at cell surface markers are of mouse origin, or if the antibodies are conjugated with biotin, the antibody-bound cells can be removed by an avidin-coated surface; or if the antibodies are conjugated to magnetic beads, the cells expressing antigens recognized by the antibodies can be removed in a magnetic field.
  • Various procedures for culturing and expanding blood-borne mesenchymal cells after isolation are also known, and various examples are set forth in the Cerami et al. '446 patent. These cells express vimentin, fibronectin, collagen I and III, which are typical markers for fibroblasts.
  • these cells do not express cytokeratin, von Willebrand's factor, desmin, laminin and smooth muscle cell .alpha. -actin, all of which are commonly used markers for epithelial, endothelial or smooth muscle cells.
  • the antigens that are typically expressed on peripheral blood leukocytes such as CD3, CD4, CD8, and CD56 are also not present on the blood-borne mesenchymal cells. These cells are also positive for CD34. These cells exhibit a unique spindle-shaped morphology which is typical for fibroblasts, but atypical for other blood-derived cells.
  • the blood-borne mesenchymal cells appear to be a distinct cell type which is different from all previously described cell populations from the blood, based on their cell size, cell surface phenotype, and morphological properties.
  • Isolated blood-borne mesenchymal cells proliferate in vitro in culture media for extended periods of time using standard culture techniques that are well known to those skilled in the art. Serum-enriched medium may be used, and growth may be further enhanced by the addition of GM-CSF which accelerates the time course over which the fibroblast-like cells dominate the culture.
  • isolated cells may be engineered to express endogenous GM-CSF to sustain their long-term growth in an autocrine fashion.
  • Continuous cell lines or clones generated in this manner may facilitate further isolation of cell surface markers and cytokines and the genes encoding therefor.
  • Long-term culture of blood-borne mesenchymal cells may be performed in tissue culture flasks, roller bottles, bioreactor systems and any culture methods known in the art. In fact, these mesenchymal cells may respond to a number of other conventional cytokines and growth factors.
  • Another illustrative procedure for producing autologous fibrocytes as described in the Quan et al. article is as follows. Draw a blood sample into heparinized tubes, dilute 2:1 with saline, and separate the buffy coat from the red cells by Ficoll Hypaque density-gradient centrifugation. The leukocyte-rich buffy coat is then removed, washed, and re-suspended in DMEM supplemented with 10%, heat-inactivated fetal bovine serum. The resulting mononuclear cells then are plated onto tissue culture ware and grown in DMEM supplemented with 10% heat- inactivated fetal calf serum.
  • Fibronectin pre-coating of the culture surfaces increases the yield and better sustains the growth of fibrocytes.
  • the non-adherent cells largely T cells
  • the remaining adherent cells cultivated for 14 days.
  • the contaminating monocytes die off, and fibrocytes appear as clusters of stellate, elongated or spindle-shaped cells that show long cellular processes.
  • fibrocytes then begin contracting into a fusiform shape.
  • Another illustrative procedure for generating PFPC is to collect a preparation of mononuclear blood cells from a patient with conventional hemapheresis devices (blood cell separators), or to obtain the mononuclear blood cells from whole blood using conventional centrifugation methods. Immediately thereafter or after a second purification step using conventional density gradient centrifugation methods, the cells are cultured in a media absent of human autologous or donor plasma for 1-7 days. It is expected that a significant population of PFPCs (more than 5%) would develop in this culture step. After re-suspension and rinsing of the non-adherent cells that contain the PFPC fraction, the final cell product may be used for therapy.
  • the PFPCs could be isolated with one or more specific antibodies using conventional laboratory methods such as immunomagnetic isolation.
  • the isolation could be either positive, i.e. by targeting the PFPCs with specific antibodies and then releasing them; or negative, i.e. by targeting the non-PFPC cell populations with specific antibodies and then discarding them.
  • the final, highly purified product may then be used for therapy.
  • PFPCs mediates tissue repair and may differentiate into fibroblasts. Particularly where injury to the tissue has occurred, PFPCs rapidly enters the site of tissue injury, where they secretes inflammatory cytokines and extra-cellular matrix (“ECM”) proteins, and promotes angiogenesis and wound contraction.
  • ECM extra-cellular matrix
  • PFPCs may contribute to the spread of excitation via functional gap junctions with cardiomyocytes, in the manner of fibroblasts.
  • a temporary conduction block is created. Over time, the extra-cellular matrix is replaced by new myocardium so that normal or at least some conduction is restored.
  • PFPCs into the atrioventricular node CAVN is particularly advantageous for moderating electrical activity in the left ventricle, although introduction of PFPCs into other electrically active areas of the heart such as the sino-atrial node ("SAN") and the conducting fibers of the ventricles is also possible. Moreover, PFPCs may be introduced into multiple sites to achieve particular beneficial results.
  • PFPCs may be administered to patients via any of a number of routes, including but not limited to epicardial or endocardial using percutaneous, intravenous, or open chest delivery techniques, as desired.
  • FIG. 1A shows one illustrative technique for administering autologous
  • PFPCs to the heart of a patient using essentially the production technique of Pilling et al.
  • Bone marrow lineage cells are isolated (block 8) from whole blood taken from the patient.
  • SAP is depleted from the plasma (block 9) so that differentiation may proceed rapidly, and the bone marrow lineage cells are incubated to produce PFPCs (block 19).
  • the concentrated PFPCs are introduced into the heart of the patient to modify electrical conduction in the heart. Examples of how the PFPCs may be introduced are shown in FIG. 2 through FIG. 5.
  • FIG. 1B shows another illustrative technique for administering autologous PFPCs to the heart of a patient using essentially the production technique of Cerami et al.
  • PFPCs and/or other bone marrow lineage cells are isolated (block 21 ) from whole blood taken from the patient, and are cultured to produce PFPCs (block 23).
  • the concentrated PFPCs are introduced into the heart of the patient to modify electrical conduction in the heart. Examples of how the PFPCs may be introduced are shown in FIG. 2 through FIG. 5.
  • FIG. 2 shows a heart 1 into which is introduced by any suitable technique a PFPCs-containing agent 2.
  • the agent 2 may be PFPCs that are simply suspended in a suitable medium such as, for example, culture medium, diluted plasma without SAP, Dulbecco's modified eagle medium ("DMEM"), and so forth.
  • the agent 2 may include PFPCs along with one or more other materials such as an injectable biopolymer, a collagen or precursor or analog or derivative thereof, a cell culture, skeletal myoblasts, stem cells, fibroblasts, any of the materials described in the '740 published application, and other such materials.
  • PFPCs may be combined with other materials to achieve multiple therapeutic effects and/or a different release kinetics for the PFPCs.
  • One suitable technique of introducing the agent 2 is by injection with a single lumen needle.
  • FIG. 3 shows a heart 3 into which is introduced by any suitable technique a multiple component agent.
  • Mixing of the components may occur outside of the tissue as they are being introduced into the tissue, or in the tissue after the components have been separately introduced into the tissue.
  • the separate introduction of the components may be achieved either by a small degree of spatial separation if they are introduced at the same time, or by sequential application if they are introduced into the same tissue site.
  • a dual-component agent 4 has a PFPCs-containing component 5 and another component 6. If desired, PFPCs may be provided in more than one of the components.
  • the components may be materials that are not suitable for mixing prior to injection, including agent precursors such as the fibrin glue precursors fibrinogen and thrombin - the thrombin converts the fibrinogen to fibrin by enzymatic action at a rate determined by the concentration of thrombin.
  • the components may be materials that are injected into the same site sequentially.
  • materials that may be mixed prior to injection to form an agent may be administered as components of the agent as shown in FIG. 3.
  • One suitable technique of introducing the agent 4 is by injection with a dual lumen needle having a mixing orifice at the distal end or separate orifices at the distal end.
  • FIG. 4 is a variation of the multiple component agent approach shown in FIG. 3, wherein the agent 12 includes a differentiation promoter 14 along with the PFPCs component 13.
  • the differentiation promoter 14 is introduced to a particular area of the heart so that any bone marrow derived lineage cells that may traffic to the site may more rapidly differentiate to participate in the healing process.
  • a differentiation promoter is a material that binds the SAP, thereby decreasing the concentration of SAP so that the local differentiation of fibrocytes is enhanced and localized fibrosis is promoted. Suitable binding materials include DNA and chromatin from dead or damaged cells.
  • FIG. 4 shows the differentiation promoter 14 as separate from the PFPCs-containing agent 13, it may be combined with the fibrocyte component 13 under some circumstances and introduced as shown in FIG. 2, for example.
  • FIG. 5 shows a heart 18 into which a PFPCs-containing agent 12 is introduced substantially as shown in FIG. 4.
  • a differentiation promoter 14 is introduced to a particular area of the heart so that any bone marrow derived lineage cells that traffic to the site differentiate to mediate the healing process.
  • a differentiation inhibitor 17 may be introduced into these areas.
  • a suitable differentiation inhibitor is SAP. This approach allows for regulation of a potentially excessive fibrotic response that could threaten pathological scaring.
  • the agent 12 and the differentiation inhibitor 17 may be introduced separately using different instruments, or may be introduced along with the agent 12 using an integrated instrument.
  • One suitable technique for introducing the agent 12 along with the differentiation inhibitor 17 is by injection with a three lumen needle in which two of the lumen which convey the PFPCs component 13 and the differentiation promoter 14 merge at the proximal end at a mixing orifice, which is surrounded at the distal end by the third lumen which conveys the differentiation inhibitor 17.
  • the fibrocyte component 13 and the differentiation promoter 14 may be combined in a single container (not shown) and injected along with the differentiation inhibitor 17 in a two lumen needle, in which the inner lumen conveys the agent 12 and the outer lumen conveys the differentiation inhibitor 17.
  • FIG. 6 illustratively shows a source 10 of material 15 and a single lumen delivery catheter 20. Delivery catheter 20 delivers material 15 to a region of a heart in a patient, such as shown for example in FIG. 11 , and is suitable for injecting the agent 2 shown in FtG. 2.
  • the delivery catheter 20 illustratively has an elongate body 22 with a proximal end portion 24, a distal end portion 28, and a lumen 32 extending therethrough between proximal and distal ports 34 and 38 located along proximal and distal end portions 24 and 26, respectively.
  • Proximal port 34 includes a proximal coupler 36 for coupling (not shown) to the source 10.
  • a needle 40 extends beyond distal tip 29 for insertion into tissue and to deliver material 15 into the tissue. Needle 40 may be fixed or axially moveable (as shown) relative to catheter 20.
  • the single lumen 32 slideably houses needle 40, which in turn includes its own delivery lumen for carrying the material 15 from the source 10 into the target tissue.
  • Lumens may be provided, for example, to house pull- wires, push-wires, filaments, fibers, or torsional members for deflecting the distal end portion 28 in-vivo; or to house guidewires, or to house leads to mapping electrodes such as may be provided at or near the tip 29 or otherwise along distal end portion 28.
  • the needle 40 may take many different forms, such as a relatively straight sharp-tip needle, or may be a hollow screw-shaped needle or other mechanism, such as to aid in anchoring at the desired location.
  • the delivery catheter 20 may be varied in many different ways to provide for delivery of material.
  • One or more needles may be provided at other places than at the tip 29, such as along the side wall of the elongate body of distal end portion 28 of the catheter 20.
  • multiple needles may be deployed along the length of the catheter 20, or a single needle may be moved to different locations along the length of the catheter 20.
  • FIG. 7 shows a system 100 having a delivery catheter 120 that injects an agent derived from separate containers 112 and 116 containing two separate components 114 and 118 of the agent, and is suitable for injecting the agent 4 shown in FIG. 3, and the agent 12 shown in FIGS. 4 and 5.
  • the two materials 114 and 118 may be precursor materials to forming the agent, wherein one or more of the precursor materials also contains the PFPCs.
  • fibrin glue for example, the combined delivery of the precursor materials, either as the separate precursor materials that are later mixed, or in combined form mixed as fibrin glue, is hence considered a fibrin glue "agent.”
  • agent means the end result, or the combination of precursor material components that lead to the resultant material, though in other regards “agent” may also include the desired resulting material itself.
  • FIG. 8 shows that the precursor materials 114 and 118 may be mixed within a lumen just before they are injected into the tissue through the needle 140 beyond tip 129 of distal end portion 128 as a mixed form of fibrin glue.
  • Other suitable substitutes may be used in order to achieve the objective of delivering two precursor materials and mixing them to form the media for injection.
  • FIG. 9 shows a schematic view of a system 200 wherein a distal end
  • Two separate and distinct needles 240 and 250 are used to deliver each of two materials 214 and 218, respectively, from sources 212 and 216, also respectively, located outside of the patient's body. In this manner, two materials are delivered separately into the tissue 202 where they mix within the tissue structure to form the agent 260.
  • systems 100 and 200 are useful for agents that include a combination of two materials, other materials may be substituted for use in such systems, and such systems may be appropriately modified for a particular material delivery.
  • autologous PFPCs may be delivered in combination with a second material according to either system 100 or 200.
  • second material may itself be a fibrin glue or other biopolymer agent, which may illustrate further multiples of sources and delivery lumens.
  • FIG. 10 shows a region of cardiac tissue 302 that includes an infarct zone 304 that is related to a reentrant conduction pathway 306 (illustrated in bolded arrows) associated with cardiac arrhythmia.
  • the distal end portion 328 of a catheter 320 is injected to the region at a location associated with the reentrant circuit 306. This is done by, for example, using a mapping electrode 330 provided at distal tip 329 and via an external mapping/monitoring system 336 coupled to proximal end portion 324 of catheter 320 outside of the body. Needle 340 is punctured into the tissue at the location, and is used to cause fibrocytes to populate the area of injection.
  • a differentiation inhibitor 17 may be injected in adjacent areas to preserve existing conduction pathways. According to this highly localized injection of the material 315 into the location across the reentrant circuit 306, the circuit is blocked by material 315 and its arrhythmic effects diminished or entirely remedied with hopeful return to sinus rhythm.
  • FIG. 13 One illustrative example that provides a circumferential pattern for delivery of material is shown in FIG. 13, and includes a delivery catheter 420 with an expandable member 430 on its distal end portion 428 and coupled to a proximal actuator 434 externally of the body. Expandable member 430 may be an inflatable balloon that is coupled via catheter 420 to actuator 434 that is a source of pressurized fluid. A plurality of needles 440 are provided along a circumferential band 436 of balloon 430.
  • the system 400 is particularly suitable to form a circumferential conduction block in a circumferential region of tissue at a location where a pulmonary vein extends from an atrium. It is to be appreciated that the conduction block formed by such a device and in similar manner may not be absolute or complete and still provide beneficial results.
  • FIG. 14 shows a deflectable tip design variation wherein catheter 460 has a distal end portion 468 with a balloon 466 that is deflectable by manipulating actuator 464.
  • Pull wire designs for example may be employed to achieve this embodiment.
  • FIG. 15 shows a variation in which a catheter 470 has a guidewire tracking mechanism via an internal lumen that rides over a guidewire 480 so that distal end portion 478 and balloon 476 may be delivered to the pulmonary vein where the guidewire 480 is seated.
  • an agent may be delivered by "point delivery” or along a predetermined pattern.
  • Point delivery may be achieved with a variety of different devices, including conventional needles, "end- hole” injectors, screw needles with multiple ports along the screw shank, and the needle devices provided herein with multiple adjacent needles intended to provide localized mixing in tissues.
  • other devices provided delivery along a predetermined pattern of tissue along the respective cardiac tissue structure (e.g. wall) beyond a single injection site as would result from such needle or end-hole devices.
  • Such patterned delivery and resulting conduction block generally provides pre-determined geometry with varied dimensions (e.g.
  • patterned delivery systems may include provision for injecting differentiation inhibitors as well as differentiation inducers.
  • the needles in each type of delivery device may be of a single lumen type or a multiple lumen type.
  • Other suitable systems include other modes for delivering the desired material, such as through walls of porous membranes forming such a circumferential band.
  • Other devices than a balloon may be used as well, such as expandable members such as cages, or other devices such as loop-shaped elongate members that may be configured with appropriate dimension to form the desired circumferential block.
  • other blocks than circumferential blocks may be made and still provide benefit.
  • other conduction blocks may be done such as similar to the "maze" procedure and using similar techniques to those previously described using ablation technology.
  • contact members such as cages, balloons, screw or needle anchors, may be used in order to anchor a delivery assembly in place so that needles or other injection or delivery members may be then extended from a position along the delivery catheter to another location adjacent to the contact member.
  • contact members may include the needles themselves, and multiple needles may be employed in a spaced fashion over a pattern for delivery, allowing for the injection and subsequent diffusion or other transport mechanisms in the tissue to close the gaps and complete the pattern as one example of an equivalent approach to continuous, uninterrupted contact of a delivery member over that pattern.
  • "contacting" a patterned region of tissue is considered contextual to the particular embodiment or application, and may be substantially continuous and uninterrupted contact in certain circumstances, or in others may have interruptions that are considered insignificant in the context of the anatomy or more general use.
  • PFPCs introduction in conjunction with ablation techniques may be practiced if desired.
  • fibrocytes may be introduced with or without other agents, in conjunction with ablation to a greater or lesser degree.
  • PFPCs may be introduced alone or along with one or more different materials to modify conduction in the myocardium.
  • highly beneficial materials include cells, peptides, biopolymers, and other fluids or preparations that interfere with intercellular junctions, such as impeding communication across or physically separating cellular gap junctions.
  • Biopolymer agents such as fibrin glue are beneficial, and typically is provided as two precursor materials that are mixed to form fibrin glue.
  • Another highly beneficial example includes an injectable material containing collagen, or a precursor or analog or derivative thereof.
  • the collagen, or precursor or analog or derivative materials thereof may include, for example, a carrier or matrix that adapts the collagen for delivery and may or may not be otherwise be retained with the collagen when implanted to the location, or may otherwise be transported or metabolized at the injection site.
  • suitable material include cells such as myoblasts, fibroblasts, stem cells, and other suitable cells that provide sufficient gap junctions with cardiac cells to form conduction block.
  • cells such as myoblasts, fibroblasts, stem cells, and other suitable cells that provide sufficient gap junctions with cardiac cells to form conduction block.
  • they may be cultured from the patient's own cells, or may be foreign to the body, such as from a regulated cell culture.
  • the delivery of these materials may be done in a highly localized manner at locations along arrhythmic pathways, or at focal sources of arrhythmia, in order to focus the conduction blocking effects in a positive manner to treat arrhythmias with localized, cellular conduction blocks.
  • PFPCs blood-borne mesenchymal cells
  • MHC major histocompatibility complex
  • Pulsing PFPCs with antigen results in modulation of T cell- mediated immunity and thus affects antigenicity.
  • the immune response may be enhanced due to the antigen presenting capability of the fibrocyte cells and stimulation of T lymphocytes.
  • the mesenchymal cells may be genetically engineered to express one or more desired gene products. The engineered cells may then be administered in vivo (e.g., either returned to the autologous host or administered to an appropriate recipient) to deliver their gene products locally or systemically.
  • precursor or analogs or derivatives thereof are further contemplated, such as for example structures that are metabolized or otherwise altered within the body to form collagen, or combination materials that react to form collagen, or material whose molecular structure varies insubstantial Iy to that of collagen such that its activity is substantially similar thereto with respect to the intended uses contemplated herein (e.g. removing or altering non-functional groups with respect to such function).
  • a group of collagen and such precursors or analogs or derivatives thereof is herein referred to as a "collagen agent.”
  • reference herein to other forms of "agents”, such as for example “polymer agent” or “fibrin glue agent” may further include the actual final product, e.g. polymer or fibrin glue, respectively, or one or more respective precursor materials delivered together or in a coordinated manner to form the resulting material.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Cell Biology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Veterinary Medicine (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Virology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Hematology (AREA)
  • Cardiology (AREA)
  • Vascular Medicine (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne des cellules désignées cellules souches sanguines à activité profibrotique (PFPC) utiles pour le traitement sans ablation de perturbations de conduction dans le coeur. Des PFPC autologues concentrées ont la capacité de modifier des voies électriques du coeur de diverses manières, comprenant la conduction par diffusion ainsi que par bloc de conduction au fur et à mesure de la formation de tissu cicatriciel suite à une sécrétion de collagène, de facteurs de croissance probiotiques, et de cytokines. Lorsque le couplage électromécanique de fibroblastes peut être induit, des PFPC peuvent également être utilisées pour créer de nouvelles voies pour normaliser la conduction du coeur à partir de voies de conduction anormales. D'autres techniques peuvent être utilisées, comprenant la production de protéines des jonctions lacunaires au moyen des PFPC afin de normaliser la voie de conduction par le couplage électromécanique avec les myocytes cardiaques existants.
PCT/US2007/009168 2006-04-19 2007-04-12 Procédé et appareil pour modifier la conduction électrique dans le coeur au moyen de cellules souches sanguines à activité profibrotique WO2007123851A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US79334006P 2006-04-19 2006-04-19
US60/793,340 2006-04-19

Publications (2)

Publication Number Publication Date
WO2007123851A2 true WO2007123851A2 (fr) 2007-11-01
WO2007123851A3 WO2007123851A3 (fr) 2008-12-24

Family

ID=38625524

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2007/009168 WO2007123851A2 (fr) 2006-04-19 2007-04-12 Procédé et appareil pour modifier la conduction électrique dans le coeur au moyen de cellules souches sanguines à activité profibrotique

Country Status (1)

Country Link
WO (1) WO2007123851A2 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050271631A1 (en) * 2002-11-29 2005-12-08 Lee Randall J Material compositions and related systems and methods for treating cardiac conditions

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050271631A1 (en) * 2002-11-29 2005-12-08 Lee Randall J Material compositions and related systems and methods for treating cardiac conditions

Also Published As

Publication number Publication date
WO2007123851A3 (fr) 2008-12-24

Similar Documents

Publication Publication Date Title
Nagaya et al. Intravenous administration of mesenchymal stem cells improves cardiac function in rats with acute myocardial infarction through angiogenesis and myogenesis
JP4562816B2 (ja) 心筋再生のための間葉幹細胞の使用法および組成物
US20040005295A1 (en) System and method for treating cardiac arrhythmias with fibroblast cells
US20200254019A1 (en) Pharmaceutical preparation
US7658951B2 (en) Method of improving cardiac function of a diseased heart
US20040002740A1 (en) System and method for forming a non-ablative cardiac conduction block
Tang et al. VEGF-A promotes cardiac stem cell engraftment and myocardial repair in the infarcted heart
Zhou et al. Regulatory T cells enhance mesenchymal stem cell survival and proliferation following autologous cotransplantation in ischemic myocardium
Taghavi et al. Autologous c‐Kit+ mesenchymal stem cell injections provide superior therapeutic benefit as compared to c‐Kit+ cardiac‐derived stem cells in a feline model of isoproterenol‐induced cardiomyopathy
WO2011022071A2 (fr) Compositions cardiaques
US20080089867A1 (en) Method of increasing retention, survival and proliferation of transplanted cells in vivo
Hansen et al. Development of a contractile cardiac fiber from pluripotent stem cell derived cardiomyocytes
KR20070026319A (ko) 골수-전구세포(ｂｍｐ) 및/또는 혈액-유래 순환전구세포(ｂｄｐ)의 심혈관 기능을 진단하는 시험관내 방법
McCully et al. Mitochondrial transplantation: the advance to therapeutic application and molecular modulation
US20100034794A1 (en) Endothelial progenitor cell compositions and neovascularization
WO2017192602A1 (fr) Utilisations d'inhibiteurs de la transition épithélio-mésenchymateuse dans la génération de cellules pacemaker
EP1972684A1 (fr) Milieu de culture de cellules souches autologues humaines et ses applications
Al-Khani et al. Mesenchymal Stem Cells for Cardiac Repair: The Clinical Perspective
WO2007123851A2 (fr) Procédé et appareil pour modifier la conduction électrique dans le coeur au moyen de cellules souches sanguines à activité profibrotique
CN110669727A (zh) 骨髓间充质干细胞膜片的制备方法及其应用
Hou et al. Establishing a new electrical conduction pathway by anastomosis of the right auricle and right ventricle assisted by mesenchymal stem cells in a canine model
US20180135017A1 (en) Composition and method of stem cells for preservation of cardiac tissue
Laberge et al. The role of microvesicles in cutaneous wound healing
ES2666351T3 (es) Composición y método de células madre para la preservación del tejido cardíaco
余川順一郎 Enhanced Device for Cell Delivery to the Myocardium: Validation in Swine Hearts

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07755439

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 07755439

Country of ref document: EP

Kind code of ref document: A2