Reversing the PAI-1-induced fibrotic immune exclusion of solid tumor by multivalent CXCR4 antagonistic nano-permeator

Fibrosis is one of the key factors that lead to the immune exclusion of solid tumors. Although degradation of fiber is a promising strategy, its application was still bottlenecked by the side effects of causing metastasis, resulting in the failure of immunotherapy. Here, we developed an antimetastatic polymer (HPA) for the delivery of chemo-drug and antifibrotic siPAI-1 to form the nano-permeator. Nano-permeator shrank after protonation and deeply penetrated into the tumor core to down-regulate the expression of PAI-1 for antifibrosis, and further promoted the sustained infiltration and activation of T cells for killing tumor cells. Moreover, metastasis after fiber elimination was prevented by multivalent CXCR4 antagonistic HPA to reduce the attraction of CXCL12 secreted by distant organs. The administration of stroma-alleviated immunotherapy increased the infiltration of CD8+ T cells to 52.5% in tumor tissues, inhibiting nearly 90% metastasis by HPA in distant organs. The nano-permeator reveals the mechanism and correlation between antifibrosis and antimetastasis and was believed to be the optimizing immunotherapy for solid fibrotic tumors.

Intervention of AMD3100 in lung tissues of rats during pulmonary oxygen intoxication / 军事医学

Objective To investigate the intervention of chemokine receptor 4(CXCR4) antagonist AMD3100 in lung tissues of rats during pulmonary oxygen intoxication.Methods Forty SD rats were randomly divided into 4 groups:normal pressure air PBS group, normal pressure air antagonist group , oxygen exposure PBS group and oxygen exposure antagonist group, each consisting of 10 animals.The last two groups were compressed to 0.23 MPa at an exponential rate of 0.1 MPa/min by pure oxygen.Pathological changes of lung tissues were observed by hematoxylin eosin stain.Changes in TNF-αand IL-1βexpression levels in the lung tissues of rats were detected by ELISA.Changes in CXCR4 expression levels were ob-served by Western blotting.Results Pathological examination indicated that edema and hemorrhage in the alveolar and pulmonary interstitial tissue of oxygen exposure antagonist group were lighter than in oxygen exposure PBS group.The levels of TNF-α, IL-1βand cleaved-caspase-3 in the lung tissues of the oxygen exposure antagonist group were lower than in oxy-gen exposure PBS group.Conclusion Blocking CXCR4 with AMD3100 can effectively alleviate lung injury during pulmo-nary oxygen intoxication.

Effect of Endogenous Bone Marrow Derived Stem Cells Induced by AMD-3100 on Expanded Ischemic Flap

The purpose of this study was to devise an expanded ischemic flap model and to investigate the role of AMD-3100 (Plerixafor, chemokine receptor 4 inhibitor) in this model by confirming its effect on mobilization of stem cells from the bone marrow. Male Sprague-Dawley rats were used as an animal research model. The mobilization of stem cells from the bone marrow was confirmed in the AMD-3100-treated group. The fractions of endothelial progenitor cells (EPC) and the vascular endothelial growth factor receptor (VEGFR) 2+ cells in the peripheral blood were increased in groups treated with AMD-3100. The expression of vascular endothelial growth factor (VEGF) was increased in response to expansion or AMD injection. The expression of stromal cell derived factor (SDF)-1 and VEGFR2 were increased only in unexpanded flap treated with AMD-3100. Treatment with AMD-3100 increased both the number and area of blood vessels. However, there were no statistically significant differences in the survival area or physiologic microcirculation in rats from the other groups. This endogenous neovascularization induced by AMD-3100 may be a result of the increase in both the area and number of vessels, as well as paracrine augmentation of the expression of VEGF and EPCs. However, the presence of a tissue expander under the flap could block the neovascularization between the flap and the recipient regardless of AMD-3100 treatment and expansion.

Effects and Significance of SDF-1/CXCR4 in Proliferation, Migration and Invasion of Colorectal Cancer Cell Line SW480 / 天津医药

Objective To discuss the influence and significance of stromal cell-derived factor 1 (SDF-1) and its specific receptor CXC chemokine receptor 4 (CXCR4) in proliferation, migration and invasion ability of SW480 colorectal cancer cells. Methods The colorectal cancer cell line SW480 in logarithmic phase was divided into four groups:control group (with no any processing), SDF-1 group (added 100μg/L SDF-1), SDF-1+1 mg/L AMD3100 mixed group (added 1 mg/L AMD3100 for 2 hours, then added 100μg/L SDF-1) and AMD3100 group (added 1 mg/L AMD3100). Immunohisto?chemistry method was used to detect the protein expression of CXCR4 in SW480 cells. The expression of CXCR4 mRNA in SW480 cells was detected by RT-PCR before and after SDF-1 and AMD3100 treatment. MTT assay and transwell chamber were used to test the changes of proliferation, migration and invasion ability of SW480 cells before and after SDF-1 and AMD3100 treatment. Results The result of immunohistochemistry showed that CXCR4 protein was expressed in SW480 cells (positive rate=80%). CXCR4 mRNA was expressed in SW480 cells. The expression of CXCR4 mRNA was up-regulat?ed by SDF-1(100μg/L), which could be inhibited by AMD3100 (1 mg/L). The proliferation activity was higher in SDF-1 group (0.847±0.039) compared to that in control group (0.624±0.011) and SDF-1+AMD3100 mixed group (0.607 ±0.016). The proliferation activity was lower in AMD3100 group (0.456 ± 0.031) than that in control group and SDF-1+AMD3100 mixed group (F=108.03, P<0.05). The number of transmembrane cells was more in SDF-1 group (98.7±5.8, 33.7±6.2) than that in control group (21.0±2.2, 6.1±2.3), SDF-1+1 mg/L AMD3100 mixed group (18.5±8.4, 8.5±2.8) and AMD3100 group (12.1±3.2, 2.1±1.0) detected by transwell chamber experiment. However, there were no statistical differences between three groups. Conclusion The biological axis SDF-1/CXCR4 can promote the proliferation, migration and invasion in colorectal cancer cell line SW480.

Bone marrow stem/progenitor cell mobilization in C57BL/6J and BALB/c mice / 한국실험동물학회지

Bone marrow (BM) has been considered as a reservoir of stem/progenitor cells which are able to differentiate into ectodermal, endodermal, and mesodermal origins in vitro as well as in vivo. Following adequate stimulation, such as granulocyte stimulating factor (G-CSF) or AMD3100, BM resident stem/progenitor cells (BMSPCs) can be mobilized to peripheral blood. Several host-related factors are known to participate in this mobilization process. In fact, a significant number of donors are resistant to G-CSF induced mobilization protocols. AMD3100 is currently used in combination with G-CSF. However, information regarding host-related factors which may influence the AMD3100 directed mobilization is extremely limited. In this study, we were to get some more knowledge on the host-related factors that affect the efficiency of AMD3100 induced mobilization by employing in vivo mobilization experiments. As a result, we found that C57BL/6J mice are more sensitive to AMD3100 but less sensitive to G-CSF which promotes the proliferation of BMSPCs. We excluded S1P as one of the host related factor which influences AMD3100 directed mobilization because pre-treatment of S1P receptor antagonist FTY720 did not inhibit BMSPC mobilization. Further in vitro experiments revealed that BALB/c mice, compared to C57BL/6J mice, have less BMSPCs which migrate in response to host related factors such as sphingosine-1-phosphate (S1P) and to CXCL12. We conclude that AMD3100-directed mobilization depends on the number of BMSPCs rather than on the host-related factors. These results suggest that the combination of AMD3100 and G-CSF is co-operative and is optimal for the mobilization of BMSPCs.

Study on effects of G-CSF and AMD3100 on proliferation, migration and adhesion of rat mesenchymal stem cells / 国际生物医学工程杂志

Objective To study the effects ofgranulocyte colony-stimulating factor(G-CSF) and AMD3100 on the proliferation,migration and adhesion of mesenchymal stem cells (MSCs).Methods The proliferation,migration and adhesion of MSCs were detected by MTT chromometry,transwell and adhesion test.Results When the concentration of G-CSF was 200 μg/L and AMD3100 was 0.5 mg/L,the proliferation and migration of MSCs were the strongest.When the concentration of G-CSF was 200 μg/L and AMD3100 was 0 mg/L,the adhesion of MSCs was the strongest.Conclusion The proliferation,migration,adhesion of MSCs are promoted by G-CSF,and inhibited by AMD3100.

Differential effects of CXCR4 antagonists on the survival and proliferation of myeloid leukemia cells in vitro

BACKGROUND: Antagonists of CXC chemokine receptor 4 (CXCR4), including AMD3100, induce peripheral mobilization of hematopoietic stem cells and have been approved for clinical use. We explored whether the CXCR4 antagonists affected the survival and proliferation of myeloid leukemia cells in vitro. METHODS: The effects of CXCR4 antagonists AMD3100 and T140 on the survival and proliferation of myeloid leukemia cell lines (U937, HL-60, MO7e, KG1a, and K562) as well as CD34+ cells obtained from patients with AML and CML were analyzed by flow cytometry by using annexin V and a colorimetric cell proliferation assay. RESULTS: AMD3100, but not T140, stimulated the proliferation of leukemia cells in vitro in a dose-dependent manner for up to 5 days (~2-fold increase at a concentration of 10-5 M), which was not abrogated by pretreatment of the cells with pertussis toxin, but was attenuated by RNAi knockdown of CXCR7 transcripts. In contrast, AMD3100 induced a marked decrease in the cell numbers after 5-7 days. AMD3100, but not T140, induced phosphorylation of MAPK p44/p42. AMD3100 increased the number and size of leukemia cell colonies and reduced cell apoptosis during the first 5-7 days of incubation, but the phenomena were reversed during the later period of incubation. CONCLUSION: The effects of CXCR4 antagonists on the proliferation of myeloid leukemia cells are not uniform. AMD3100, but not T140, exerts dual effects, initially enhancing and subsequently inhibiting the survival and proliferation of the cells in vitro.

Effect of AMD3100 on the proliferation and angiogenesis of AsPC-1 cells / 中华胰腺病杂志

Objective To investigate the effects of blockade on non-peptide specific SDF-1/CXCR4 receptor ligand system with AMD3100 on the proliferation and angiogenesis of human pancreatic cancer cells AsPC-1. Methods AsPC-1 was divided into control group, SDF-1α group, group, SDF-1α + AMD3100 group. MTT test was performed to determine the proliferative level of AsPC-1 cells. Vascular endothelial growth factor (VEGF) was detected with Western blotting assay. Immunohistochemistry was used to detect the microvessel density (MVD) in subcutaneous xenografts of AsPC 1 of nude mice model, which was intratumorally and peritumorally injected with AMD3100. Results SDF-1α could induce the proliferation of AsPC-1(1.430 ±0. 122 vs 1. 002 ± 0. 001, P <0. 05). While the proliferative effect induced by SDF-1α could be inhibited by AMD3100 (0.983 ±0. 068vs 1.430 ± 0. 122, P <0.05). SDF-1α could induce the expression of VEGF (0. 565 ± 0. 047 vs 0. 439 ± 0.034, P < 0.05). While the protein expression of VEGF induced by SDF-1α on AsPC-1 cells was inhibited by AMD3100 (0. 450 ± 0. 071 vs 0. 565 ± 0. 04, P <0. 05). The growth and angiogenesis of subcutaneous xenografts of nude mice model were inhibited by AMD3100; the tumor inhibitory rate was 59. 5% at 24th day. The MVD of xenografts was significantly decreased (28.56 ± 6.94 vs 98.75 ± 20. 60, P < 0. 01 ). Conclusions AMD3100 could inhibit the proliferation and angiogenesis of AsPC-1 cells both in vitro and in vivo.

The CXCR4 Antagonist AMD3100 Has Dual Effects on Survival and Proliferation of Myeloma Cells In Vitro / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: AMD3100, an antagonist of the CXCR4 chemokine receptor is soon to be used clinically for the peripheral mobilization of hematopoietic stem cells (HSCs) in patients with multiple myeloma. AMD3100 has been shown to activate a G protein coupled with CXCR4 and thus acts as a partial CXCR4 agonist in vitro. Thus, we explored whether AMD3100 affected the survival and proliferation of myeloma cells in vitro. MATERIALS AND METHODS: The effects of AMD3100 on survival and proliferation of two myeloma cell lines (RPMI8226 and U266) as well as CD138+ cells obtained from several patients with multiple myeloma were analyzed by flow cytometry using annexin V and a colorimetric cell proliferation assay (CCK-8 assay). RESULTS: AMD3100, but not T140, another CXCR4 antagonist, stimulated the proliferation of myeloma cell lines and CD138+ primary human myeloma cells (-2-fold increase) in a dose-dependent manner in serum-free culture for up to 5 days, which was inhibited by pretreating the cells with pertussis toxin. AMD3100 enhanced the proliferation of U266 cells induced by interleukin-6 and partially reversed AG490-mediated growth inhibition and apoptosis induced by serum deprivation in RPMI8226 cells. AMD3100 induced the phosphorylation of Akt and MAPK p44/p42 in U266 cells and MAPK p44/p42 in RPMI8226 cells. In contrast, AMD3100 markedly increased the cell apoptosis and reduced the number of RPMI8226 cells after 5 to 7 days of culture under serum-free conditions. CONCLUSION: AMD3100 exerts dual effects, initially enhancing and subsequently inhibiting the survival and proliferation of myeloma cells, signaling via CXCR4 in vitro.