tacrolimus and plerixafor

The combination of AMD3100 and low-dose FK506 has been shown to accelerate wound healing in vivo. Although AMD3100 is known to work by releasing hematopoietic stem cells into circulation, the mechanism of FK506 in this setting has remained unknown. In this study, we investigated the activities of FK506 in human cells and a diabetic-rat wound model using a non-immunosuppressive FK506 analog named FKVP. While FKVP was incapable of inhibiting calcineurin, wound-healing enhancement with AMD3100 was unaffected. Further study showed that both FK506 and FKVP activate BMP signaling in multiple cell types through FKBP12 antagonism. Furthermore, selective inhibition of BMP signaling abolished stem cell recruitment and wound-healing enhancement by combination treatment. These results shed new light on the mechanism of action of FK506 in acceleration of wound healing, and raise the possibility that less toxic FKBP ligands such as FKVP can replace FK506 for the treatment of chronic wounds.

Topics: Animals; Benzylamines; Bone Morphogenetic Proteins; Cyclams; Diabetes Mellitus, Type 2; Disease Models, Animal; Drug Synergism; Female; Gene Knockout Techniques; Heterocyclic Compounds; Humans; Jurkat Cells; Ligands; Peptides, Cyclic; Phosphorylation; Rats; Receptors, CXCR4; Signal Transduction; Smad1 Protein; Smad5 Protein; Tacrolimus; Tacrolimus Binding Protein 1A; Wound Healing

Adhesions are a very common complication in the abdominal surgery. Animal studies and human trials have evaluated strategies designed to reduce and prevent postsurgical adhesions but few have an evidence base that justifies routine use. A strategy to prevent adhesions effectively remains an urgent need. We studied a reproducible model of intra-peritoneal adhesion formation in rats using laparotomy with several peritoneal sutures to produce the adhesions. Here we show that entraining endogenous stem cells into injury sites using the combined effect of AMD3100 and low-dose FK-506 (AF) can reduce the adhesion score significantly and abolish peritoneal adhesions in 45% of animals in a rat model of severe postsurgical intra-abdominal adhesions, compared with saline controls. Searching for mechanisms, we found AF treatment dramatically increased SDF-1 expressing cells, HGF expressing Ym1+ M2 macrophages and CD133+ stem cells in the injury sites of peritoneal surface at day 5 post-operation. Our results demonstrate that medically induced recruitment of autologous stem cells using AF significantly reduced postsurgical intra-abdominal adhesions. These findings suggest a novel effective therapeutic approach to preventing adhesions in patients.

Topics: Abdomen; AC133 Antigen; Animals; Benzylamines; Case-Control Studies; Chemokine CXCL12; Cyclams; Disease Models, Animal; Hepatocyte Growth Factor; Heterocyclic Compounds; Laparotomy; Macrophages; Peritoneum; Rats; Rats, Inbred Lew; Stem Cells; Tacrolimus; Tissue Adhesions

Rapid regeneration of the remnant liver is critical for preventing liver failure and promoting recovery after extensive liver resection. Numerous studies have demonstrated the involvement of bone marrow-derived stem cells in liver regeneration and the potential benefits of bone marrow stem cell therapy. To avoid the preparation of stem cells, we proposed in this study to mobilize endogenous bone marrow stem cells pharmacologically with a combination of AMD3100 (A), an antagonist of CXCR4 and low-dose FK506 (F). Here we show that AF combination therapy significantly increased lineage negative (Lin-) CD34+ and Lin-CD133+ stem cells in peripheral blood and enhanced recruitment of CD133+ cells into the remnant liver in a rat model of 85% partial hepatectomy. Recruiting CD133+ stem cells in the remnant liver was associated with increased proliferation of hepatic oval cells and paralleled the increased SDF-1, CXCR4 and HGF expression. Importantly, AF combination therapy increased the number of Ki67 positive hepatocytes and BrdU incorporation in the remnant liver and improved serum levels of albumin. Our results demonstrate that pharmacological mobilization of endogenous bone marrow stem cells with AF combination therapy can enhance endogenous stem cell mobilization to promote liver regeneration and improve liver function after extensive hepatectomy.

Topics: AC133 Antigen; Alanine Transaminase; Animals; Antigens, CD34; Aspartate Aminotransferases; Benzylamines; Bone Marrow Cells; Calcineurin Inhibitors; Cell Proliferation; Chemokine CXCL12; Cyclams; Drug Therapy, Combination; Female; Hematopoietic Stem Cell Mobilization; Hepatectomy; Hepatocyte Growth Factor; Heterocyclic Compounds; Liver Regeneration; Male; Rats; Rats, Inbred Lew; Receptors, CXCR4; Serum Albumin; Stem Cells; Tacrolimus; Treatment Outcome

Transplantation is now lifesaving therapy for patients with end-stage organ failure but requires lifelong immunosuppression with resultant morbidity. Current immunosuppressive strategies inhibit T cell activation and prevent donor-recipient engagement. Therefore, it is not surprising that few host cells are demonstrated in donor grafts. However, our recent small animal studies found large numbers of recipient stem cells present after transplantation and pharmacological mobilization, resulting in a chimeric, repopulated organ. We now confirm these findings in a well-characterized large animal preclinical model. Here, we show that AMD3100 and FK506 mobilization of endogenous stem cells immediately post kidney transplantation combined with repeat therapy at 1, 2, and 3 months led to drug-free long-term survival in maximally immunologically mismatched swine. Three long-term recipients have stable chimeric transplants, preserved antidonor skin graft responses, and normal serum creatinine levels despite withdrawal of all medication for 3 years.

Topics: Allografts; Animals; Anti-HIV Agents; Benzylamines; Calcineurin Inhibitors; Cyclams; Graft Rejection; Graft Survival; Hematopoietic Stem Cell Mobilization; Heterocyclic Compounds; Kidney Failure, Chronic; Kidney Transplantation; Peripheral Blood Stem Cell Transplantation; Skin Transplantation; Swine; Swine, Miniature; Tacrolimus; Transplantation Chimera; Transplantation Tolerance

Transplant tolerance allowing the elimination of lifelong immunosuppression has been the goal of research for 60 years. The induction of mixed chimerism has shown promise and has been extended successfully to large animals and to the clinic; however, it remains cumbersome and requires heavy early immunosuppression. In this study, we reported that four injections of AMD3100, a CXCR4 antagonist, plus eight injections of low-dose FK506 (0.05 mg/kg per day) in the first week after kidney transplantation extended survival, but death from renal failure occurred at 30-90 days. Repeating the same course of AMD3100 and FK506 at 1, 2 and 3 mo after transplant resulted in 92% allograft acceptance (n = 12) at 7 mo, normal kidney function and histology with no further treatment. Transplant acceptance was associated with the influx of host stem cells, resulting in a hybrid kidney and a modulated host immune response. Confirmation of these results could initiate a paradigm shift in posttransplant therapy.

Topics: Allografts; Animals; Animals, Genetically Modified; Anti-HIV Agents; Benzylamines; Calcineurin Inhibitors; Cyclams; Glomerular Filtration Rate; Graft Rejection; Graft Survival; Hematopoietic Stem Cell Mobilization; Heterocyclic Compounds; Kidney Failure, Chronic; Kidney Function Tests; Kidney Transplantation; Peripheral Blood Stem Cell Transplantation; Rats; Rats, Inbred Lew; Tacrolimus; Transplantation Chimera; Transplantation Tolerance

Stem cell therapy has shown promise in treating a variety of pathologies including skin wounds, but practical applications remain elusive. Here, we demonstrate that endogenous stem cell mobilization produced by AMD3100 and low-dose tacrolimus is able to reduce by 25% the time of complete healing of full-thickness wounds created by surgical excision. Equally important, healing was accompanied by reduced scar formation and regeneration of hair follicles. Searching for mechanisms, we found that AMD3100 combined with low-dose tacrolimus mobilized increased number of lineage-negative c-Kit+, CD34+, and CD133+ stem cells. Low-dose tacrolimus also increased the number of SDF-1-bearing macrophages in the wound sites amplifying the "pull" of mobilized stem cells into the wound. Lineage tracing demonstrated the critical role of CD133 stem cells in enhanced capillary and hair follicle neogenesis, contributing to more rapid and perfect healing. Our findings offer a significant therapeutic approach to wound healing and tissue regeneration.

Topics: Animals; Benzylamines; Cell Lineage; Cicatrix; Cyclams; Disease Models, Animal; Drug Synergism; Hair Follicle; Hematopoietic Stem Cell Mobilization; Heterocyclic Compounds; Immunosuppressive Agents; Mesenchymal Stem Cells; Mice, Inbred C57BL; Mice, Knockout; Rats, Inbred Strains; Receptors, CXCR4; Regenerative Medicine; Skin; Tacrolimus; Wound Healing

The aim of our study was to elucidate the effect of tacrolimus (FK506) and of C-X-C chemokine receptor type 4 (CXCR4), which is a receptor specific to the stromal cell-derived factor-1α (SDF‑1α), on growth and metastasis of hepatocellular carcinoma (HCC). Following treatment with different concentrations of FK506, AMD3100 or normal saline (NS), the proliferation of Morris rat hepatoma 3924A (MH3924A) cells was measured by the MTT assay, the expression of CXCR4 was analyzed with immunohistochemistry, and the morphological changes and the invasiveness of cells were studied with a transwell assay and under a scanning electron microscope, respectively. In addition, August Copenhagen Irish rat models implanted with tumor were used to examine the pathological changes and invasiveness of tumor in vivo, the expression of CXCR4 in tumor tissues and the expression of SDF‑1α in the adjacent tissues to the HCC ones, using immunohistochemistry. In vitro, FK506 (100‑1,000 µg/l) significantly promoted the proliferation of MH3924A cells (P<0.01), and increased the expression of CXCR4 in MH3924A cells, albeit with no significance (P>0.05). By contrast, AMD3100 had no effect on the proliferation of MH3924A cells, but significantly reduced the expression of CXCR4 (P<0.05). The invasiveness of MH3924A cells was significantly (P<0.01) enhanced following treatment with FK506, SDF‑1α, FK506 + AMD3100, FK506 + SDF‑1α or FK506 + AMD3100 + SDF‑1α. In vivo, tumor weight (P=0.041), lymph node metastasis (P=0.002), the number of pulmonary nodules (P=0.012), the expression of CXCR4 in tumor tissues (P=0.048) and that of SDF‑1α in adjacent tissues (P=0.026) were significantly different between the FK506-treated and the NS group. Our results suggest that FK506 promotes the proliferation of MH3924A cells and the expression of CXCR4 and SDF‑1α in vivo. Therefore, inhibiting the formation of the CXCR4/SDF‑1α complex may partly reduce the promoting effect of FK506 on HCC.

Topics: Animals; Benzylamines; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chemokine CXCL12; Cyclams; Disease Models, Animal; Heterocyclic Compounds; Immunohistochemistry; Immunosuppressive Agents; Liver Neoplasms; Lymphatic Metastasis; Rats; Receptors, CXCR4; Tacrolimus

Novel therapeutic tools to accelerate wound healing would have a major impact on the overall burden of skin disease. Lin et al. demonstrate in mice that endogenous bone marrow stem cell mobilization, produced by a pharmacologic combination of AMD3100 and tacrolimus, leads to faster and better-quality wound healing, findings that have exciting potential for clinical translation.

Topics: Animals; Benzylamines; Cyclams; Hematopoietic Stem Cell Mobilization; Heterocyclic Compounds; Mesenchymal Stem Cells; Skin; Tacrolimus

Topics: Adult Stem Cells; Benzylamines; Cyclams; Forehead; Hematopoietic Stem Cell Mobilization; Heterocyclic Compounds; Humans; Immunosuppressive Agents; Receptors, CXCR4; Regenerative Medicine; Tacrolimus; Wound Healing