In Vivo Tumorigenicity Assays Using Subcutaneous and Orthotopic Xenografts in Immunodeficient Mice

Abstract Somatic mutations in ACVR1, encoding the serine/threonine kinase ALK2 receptor, are found in a quarter of children with the currently incurable brain tumour diffuse intrinsic pontine glioma (DIPG). Treatment of ACVR1-mutant DIPG patient-derived models with multiple inhibitor chemotypes leads to a reduction in cell viability in vitro and extended survival in orthotopic xenografts in vivo, though there are currently no specific ACVR1 inhibitors licensed for DIPG. Using an Artificial Intelligence-based platform to search for approved compounds which could be used to treat ACVR1-mutant DIPG, the combination of vandetanib and everolimus was identified as a possible therapeutic approach. Vandetanib, an approved inhibitor of VEGFR/RET/EGFR, was found to target ACVR1 (Kd=150nM) and reduce DIPG cell viability in vitro, but has been trialed in DIPG patients with limited success, in part due to an inability to cross the blood-brain-barrier. In addition to mTOR, everolimus inhibits both ABCG2 (BCRP) and ABCB1 (P-gp) transporter, and was synergistic in DIPG cells when combined with vandetanib in vitro. This combination is well-tolerated in vivo, and significantly extended survival and reduced tumour burden in an orthotopic ACVR1-mutant patient-derived DIPG xenograft model. Based on these preclinical data, three patients with ACVR1-mutant DIPG were treated with vandetanib and everolimus. These cases may inform on the dosing and the toxicity profile of this combination for future clinical studies. This bench-to-bedside approach represents a rapidly translatable therapeutic strategy in children with ACVR1 mutant DIPG.

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Inhibition of Glycolysis Suppresses Cell Proliferation and Tumor Progression In Vivo: Perspectives for Chronotherapy

International Journal of Molecular Sciences ◽

10.3390/ijms22094390 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4390

Author(s):

Jana Horváthová ◽

Roman Moravčík ◽

Miroslava Matúšková ◽

Vladimír Šišovský ◽

Andrej Boháč ◽

...

Keyword(s):

Cell Proliferation ◽

Tumor Progression ◽

Umbilical Vein ◽

Cell Metabolism ◽

High Rate ◽

Ki 67 ◽

Immunodeficient Mice ◽

Inhibition Of Glycolysis

A high rate of glycolysis is considered a hallmark of tumor progression and is caused by overexpression of the enzyme 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3). Therefore, we analyzed the possibility of inhibiting tumor and endothelial cell metabolism through the inhibition of PFKFB3 by a small molecule, (E)-1-(pyridin-4-yl)-3-(quinolin-2-yl)prop-2-en-1-one (PFK15), as a promising therapy. The effects of PFK15 on cell proliferation and apoptosis were analyzed on human umbilical vein endothelial cells (HUVEC) and the human colorectal adenocarcinoma cell line DLD1 through cytotoxicity and proliferation assays, flow cytometry, and western blotting. The results showed that PFK15 inhibited the proliferation of both cell types and induced apoptosis with decreasing the Bcl-2/Bax ratio. On the basis of the results obtained from in vitro experiments, we performed a study on immunodeficient mice implanted with DLD1 cells. We found a reduced tumor mass after morning PFK15 treatment but not after evening treatment, suggesting circadian control of underlying processes. The reduction in tumor size was related to decreased expression of Ki-67, a marker of cell proliferation. We conclude that inhibition of glycolysis can represent a promising therapeutic strategy for cancer treatment and its efficiency is circadian dependent.

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Immunocompatibility of a new dual modality contrast agent based on radiolabeled iron-oxide nanoparticles

Scientific Reports ◽

10.1038/s41598-021-89117-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Maria-Argyro Karageorgou ◽

Dimosthenis Stamopoulos

Keyword(s):

Complete Blood Count ◽

Morphological Characteristics ◽

Mononuclear Phagocyte ◽

Dual Modality ◽

Immunodeficient Mice ◽

Force Microscopy ◽

Magnetic Resonance Imaging Mri ◽

Diagnostic Applications

AbstractRadiolabeled magnetic nanoparticles are promising candidates as dual-modality-contrast-agents (DMCA) for diagnostic applications. The immunocompatibility of a new DMCA is a prerequisite for subsequent in vivo applications. Here, a new DMCA, namely Fe3O4 nanoparticles radiolabeled with 68Ga, is subjected to immunocompatibility tests both in vitro and in vivo. The in vitro immunocompatibility of the DMCA relied on incubation with donated human WBCs and PLTs (five healthy individuals). Optical microscopy (OM) and atomic force microscopy (AFM) were employed for the investigation of the morphological characteristics of WBCs and PLTs. A standard hematology analyzer (HA) provided information on complete blood count. The in vivo immunocompatibility of the DMCA was assessed through its biodistribution among the basic organs of the mononuclear phagocyte system in normal and immunodeficient mice (nine in each group). In addition, Magnetic Resonance Imaging (MRI) data were acquired in normal mice (three). The combined OM, AFM and HA in vitro data showed that although the DMCA promoted noticeable activation of WBCs and PLTs, neither degradation nor clustering were observed. The in vivo data showed no difference of the DMCA biodistribution between the normal and immunodeficient mice, while the MRI data prove the efficacy of the particular DMCA when compared to the non-radiolabeled, parent CA. The combined in vitro and in vivo data prove that the particular DMCA is a promising candidate for future in vivo applications.

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RGB-Marking to Identify Patterns of Selection and Neutral Evolution in Human Osteosarcoma Models

Cancers ◽

10.3390/cancers13092003 ◽

2021 ◽

Vol 13 (9) ◽

pp. 2003

Author(s):

Stefano Gambera ◽

Ana Patiño-Garcia ◽

Arantzazu Alfranca ◽

Javier Garcia-Castro

Keyword(s):

Clonal Selection ◽

Clonal Evolution ◽

Phylogenetic Reconstruction ◽

Experimental Models ◽

Neutral Evolution ◽

Immunodeficient Mice ◽

Nsg Mice ◽

Clonal Development ◽

Clonal Composition

Osteosarcoma (OS) is a highly aggressive tumor characterized by malignant cells producing pathologic bone; the disease presents a natural tendency to metastasize. Genetic studies indicate that the OS genome is extremely complex, presenting signs of macro-evolution, and linear and branched patterns of clonal development. However, those studies were based on the phylogenetic reconstruction of next-generation sequencing (NGS) data, which present important limitations. Thus, testing clonal evolution in experimental models could be useful for validating this hypothesis. In the present study, lentiviral LeGO-vectors were employed to generate colorimetric red, green, blue (RGB)-marking in murine, canine, and human OS. With this strategy, we studied tumor heterogeneity and the clonal dynamics occurring in vivo in immunodeficient NOD.Cg-Prkdcscid-Il2rgtm1Wjl/SzJ (NSG) mice. Based on colorimetric label, tumor clonal composition was analyzed by confocal microscopy, flow cytometry, and different types of supervised and unsupervised clonal analyses. With this approach, we observed a consistent reduction in the clonal composition of RGB-marked tumors and identified evident clonal selection at the first passage in immunodeficient mice. Furthermore, we also demonstrated that OS could follow a neutral model of growth, where the disease is defined by the coexistence of different tumor sub-clones. Our study demonstrates the importance of rigorous testing of the selective forces in commonly used experimental models.

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Antigenic and Genotypic Similarity between Primary Glioblastomas and Their Derived Neurospheres

Journal of Oncology ◽

10.1155/2011/314962 ◽

2011 ◽

Vol 2011 ◽

pp. 1-16 ◽

Cited By ~ 14

Author(s):

Valentina Caldera ◽

Marta Mellai ◽

Laura Annovazzi ◽

Angela Piazzi ◽

Michele Lanotte ◽

...

Keyword(s):

Stem Cells ◽

Genetic Alterations ◽

Tumor Stem Cells ◽

Primary Tumors ◽

Immunodeficient Mice ◽

Brain Tumor Stem Cells ◽

Tumor Phenotype ◽

Fetal Bovine ◽

Primary Glioblastomas

Formation of neurospheres (NS) in cultures of glioblastomas (GBMs), with self-renewal, clonogenic capacities, and tumorigenicity following transplantation into immunodeficient mice, may denounce the existence of brain tumor stem cells (BTSCs) in vivo. In sixteen cell lines from resected primary glioblastomas, NS showed the same genetic alterations as primary tumors and the expression of stemness antigens. Adherent cells (AC), after adding 10% of fetal bovine serum (FBS) to the culture, were genetically different from NS and prevailingly expressed differentiation antigens. NS developed from a highly malignant tumor phenotype with proliferation, circumscribed necrosis, and high vessel density. Beside originating from transformed neural stem cells (NSCs), BTSCs may be contained within or correspond to dedifferentiated cells after mutation accumulation, which reacquire the expression of stemness antigens.

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Proliferation of human malignant hematopoietic cells in immunodeficient mice: suppression by antibody to pluripotent K-562 leukemia cells involves direct cytolysis and effector cells

Blood ◽

10.1182/blood.v61.6.1045.1045 ◽

1983 ◽

Vol 61 (6) ◽

pp. 1045-1053 ◽

Cited By ~ 4

Author(s):

BB Lozzio ◽

EA Machado ◽

J Mitchell ◽

CB Lozzio ◽

CJ Wust ◽

...

Keyword(s):

Gamma Globulin ◽

Hematopoietic Cells ◽

Leukemia Cells ◽

Control Group ◽

Fab Fragment ◽

Lymphoma Cells ◽

Effector Cells ◽

Immunodeficient Mice ◽

Immune Gamma Globulin

Abstract Six human hematopoetic cell lines were successfully heterotransplanted into athymic (nude) and asplenic-athymic (lasat) neonatal mice. The tumors arising from leukemia and lymphoma cells could then be serially transplanted into adult nude mice. Seven days after the fourth serial mouse passage, each mouse was treated with goat immune gamma globulin against K-562 cells. One control group was treated similarly, but with nonimmune (normal) gamma globulin, while another control group was not treated. The goat gamma globulin was not toxic for nude and lasat mice, and the immune, but not the normal, gamma globulin suppressed local subcutaneous growth of myelosarcomas, lymphosarcomas, and Burkitt lymphoma cells. On the other hand, the growth of lung, breast, and prostatic carcinomas and a melanoma of human origin were not altered by the immune gamma globulin. Since suppression of cell growth occurred equally well in decomplemented mice, a complement-mediated cytotoxicity apparently cannot be considered as responsible for the abrogation. The Fab fragment of the immunoglobulin did not suppress the growth of the myelosarcomas. We conclude that antibody suppression of the in vivo proliferation was specific for malignant hematopoietic cells and that the Fc portion of IgG is necessary for in vivo cytolysis of leukemia cells. The most probable mechanisms are direct antibody cytolysis and antibody-dependent macrophage-mediated cytotoxicity.

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Immuno– enhancement effect of the Nam Dia Long decoction on immuno suppressed mice induced by cyclophosphamide

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v1i6.617 ◽

2018 ◽

Vol 1 (6) ◽

pp. 68-75

Author(s):

Nuong Thi My Nguyen ◽

Ngoc Thi Nhu Bui ◽

Mai Tuyet Au ◽

Hiep Minh Dinh

Keyword(s):

T Cells ◽

Cd8 T Cells ◽

Enhancement Effect ◽

Therapeutic Activity ◽

Immunodeficient Mice ◽

Black Bean ◽

Traditional Remedy ◽

Scientific Report ◽

Nam Dia Long

Nam Dia Long (NDL) formula which is composed of earthworm, black bean, mung bean and sweet leaf has been empirically used for the treatment of cancer, arthritis, epilepsy… However, there is no scientific report about the therapeutic activity of this traditional remedy. The aim of this work was to study on the the in vivo immunostimulating effect of NDL formula. In cyclophosphamide - induced immunodeficient mice, NDL powder at two doses of 1.2 g/kg and 2.4 g/kg attenuated the decrease of body weight, increased relatively the weights of spleen and thymus. These two doses also rised 42–44 % of leukocytes, 34 – 43 % of CD4 T cells and 35 – 46% of CD8 T cells in comparison with the pathological control. Therefore, NDL formula showed in vivo immunostimulating effect and has the potential to be developed as an adjuvant drug in cancer chemotherapy.

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Guiding T lymphopoiesis from pluripotent stem cells by defined transcription factors

10.1101/660977 ◽

2019 ◽

Author(s):

Rongqun Guo ◽

Fangxiao Hu ◽

Qitong Weng ◽

Cui Lv ◽

Hongling Wu ◽

...

Keyword(s):

Stem Cells ◽

Pluripotent Stem Cells ◽

Time Window ◽

Immune Surveillance ◽

Cell Stage ◽

Immunodeficient Mice ◽

Cell Transcriptome ◽

T Lymphopoiesis

ABSTRACTAchievement of immunocompetent and therapeutic T lymphopoiesis from pluripotent stem cells is a central aim in T cell regenerative medicine. To date, preferentially regenerating T lymphopoiesis in vivo from pluripotent stem cells (PSC) remains a practical challenge. Here we documented that synergistic and transient expression of Runx1 and Hoxa9 restricted in the time window of endothelial to hematopoietic transition and hematopoietic maturation stages induced in vitro from PSC (iR9-PSC) preferentially generated engraftable hematopoietic progenitors capable of homing to thymus and developing into mature T (iT) cells in primary and secondary immunodeficient recipients. Single-cell transcriptome and functional analyses illustrated the cellular trajectory of T lineage induction from PSC, unveiling the T-lineage specification determined at as early as hemogenic endothelial cell stage and identifying the bona fide pre-thymic progenitors. The iT cells distributed normally in central and peripheral lymphoid organs and exhibited abundant TCRαβ repertoire. The regenerative T lymphopoiesis rescued the immune-surveillance ability in immunodeficient mice. Furthermore, gene-edited iR9-PSC produced tumor-specific-T cells in vivo that effectively eradicated tumor cells. This study provides insight into universal generation of functional and therapeutic T lymphopoiesis from the unlimited and editable PSC source.

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Does Fingolimod have anticancer effects? A meta-analysis and systematic review based on experimental animal models of various cancers

10.21203/rs.3.rs-121127/v1 ◽

2020 ◽

Author(s):

Yuze Ge ◽

Ping Zhou ◽

Lu Kong

Keyword(s):

Renal Cell Carcinoma ◽

Animal Models ◽

Cell Carcinoma ◽

Renal Cell ◽

Meta Analysis ◽

Signal Pathways ◽

Bias Analysis ◽

Immunodeficient Mice ◽

Anticancer Effects

Abstract Background: Numerous studies have explored the anticancer effect of FTY720 (Fingolimod) in animal models, a sphingosine-1-phosphate (S1P) receptor antagonist and an immunosuppressant, but little clinical evidence guides the use of FTY720 in cancer patients.Methods: Strictly, only related published articles about the treatment with FTY720 for various cancers in vivo from January 1998 to January 2020 were selected from PubMed, Web of Science, Ovid, Embase, CNKI and Cochrane databases, and which were qualified. We acquired agreement through discussion. Then, we conducted meta-analysis, subgroups analysis, publication bias analysis and sensitivity analysis based on selected studies. In the last two sections, we summaried and compared side effects, drug combination effects and molecular pathways from selected studies.Results: In the 31 articles included from 2002 to 2019, FTY720 was found to reduce tumor volume (SMD =-2.58, 95% CI: -3.42, -1.75, Z = 6.09, P = 0.000), tumor weight (SMD = -3.69, 95% CI: -5.17, -2.21, Z = 4.88, P = 0.000) and body weight (SMD = -0.86, 95% CI: -1.61, -0.11, Z = 2.23, P = 0.025) in 14 types of cancer. Relevant frequent signal pathways include the Akt pathway, S1PRs-Caspase pathway and the STAT3-PP2A pathway. FTY720 has significant independent or in combination anticancer effects and a lower toxicity in renal cell carcinoma and neuroblastoma mice models. However, it should be noted that FTY720 achieved a significant therapeutic effect in immunodeficient mice, not in immunecompetent mice. Also, the dosage-safety of FTY720 alone in clinical use is a noteworthy issue. In mouse models, the mechanism of the FTY720 treatment of tumors lies in inducing the tumor cells apoptosis through important signaling moleculars.Conclusions: FTY720 alone or in combination exerted significant anti-tumor effects for neuroblastoma and renal cell carcinoma, however not for melanoma. Due to insufficient evidence, more specific studies of FTY720 only and in combination included in immunity, inflammation and melanoma should be carried out in the future preclinical and clinical studies.

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Exosome-mediated hematopoietic rejuvenation in a humanized mouse model indicate potential for cancer immunotherapy

10.1101/2020.08.02.233015 ◽

2020 ◽

Author(s):

Steven J. Greco ◽

Seda Ayer ◽

Khadidiatou Guiro ◽

Garima Sinha ◽

Robert J. Donnelly ◽

...

Keyword(s):

Breast Cancer Cells ◽

Preventive Measure ◽

Economic Costs ◽

Humanized Mouse ◽

Humanized Mouse Model ◽

Immunodeficient Mice ◽

Bidirectional Communication ◽

Exosomal Mirnas ◽

Mobilized Peripheral Blood

AbstractAging is associated with increased morbidity and high economic costs due to a burdened healthcare system and decreased workforce. Parabiotic animal models indicated that secretome from young cells can restore aged tissue functions. We used a heterochronic co-culture system with young and aged mobilized peripheral blood (MPB) or umbilical cord blood (UCB) and showed hematopoietic restoration, independent of allogeneic difference. Bidirectional communication between the aged and young cells influenced the miRNA cargo of exosomes, resulting in partial reprograming of the aged cells. The restored cells enhanced hematopoiesis (e.g., increased lymphoid:myeloid ratio) in immunodeficient mice bearing autologous aged hematopoietic system. Four exosomal miRNAs targeting PAX and PPMIF were partly responsible for the hematopoietic rejuvenation. Notably, increased natural killer (NK) cells within the restored cells eliminated dormant breast cancer cells in vivo. The findings could be developed as preventive measure and treatment for sustained immune/hematopoietic competence with potential for immunotherapy.

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