Use of a Human Artificial Chromosome for Delivering Trophic Factors in a Rodent Model of Amyotrophic Lateral Sclerosis

Watanabe, Yasuhiro; Kazuki, Yasuhiro; Kazuki, Kanako; Ebiki, Mitsutaka; Nakanishi, Mami; Nakamura, Kazuomi; Yamakawa, Masafumi; Hosokawa, Hiroyuki; Ohbayashi, Tetsuya; Oshimura, Mitsuo; Nakashima, Kenichiro

doi:10.1038/mtna.2015.28

Cited by 20 publications

(27 citation statements)

References 22 publications

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“…Intrathecal delivery of Riluzole has been investigated in both Gottingen minipigs and canine models [52,53] with both studies demonstrating higher Riluzole levels in the CNS while limiting the systemic dose that may lead to off-target side-effects. Additionally, gene therapy and therapies involving trophic factors to stimulate dying neurons [54,55] have shown promise in rodent models. Other animal studies have also shown that human stem cells administered intrathecally delay the onset of symptoms and prolong survival in ALS transgenic mice [56].…”

Section: Potential Implications For Intrathecal Als Therapeuticsmentioning

confidence: 99%

Non-invasive MRI quantification of cerebrospinal fluid dynamics in amyotrophic lateral sclerosis patients

Sass

Khani

Romm

et al. 2020

Fluids Barriers CNS

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Background: Developing novel therapeutic agents to treat amyotrophic lateral sclerosis (ALS) has been difficult due to multifactorial pathophysiologic processes at work. Intrathecal drug administration shows promise due to close proximity of cerebrospinal fluid (CSF) to affected tissues. Development of effective intrathecal pharmaceuticals will rely on accurate models of how drugs are dispersed in the CSF. Therefore, a method to quantify these dynamics and a characterization of differences across disease states is needed. Methods: Complete intrathecal 3D CSF geometry and CSF flow velocities at six axial locations in the spinal canal were collected by T2-weighted and phase-contrast MRI, respectively. Scans were completed for eight people with ALS and ten healthy controls. Manual segmentation of the spinal subarachnoid space was performed and coupled with an interpolated model of CSF flow within the spinal canal. Geometric and hydrodynamic parameters were then generated at 1 mm slice intervals along the entire spine. Temporal analysis of the waveform spectral content and feature points was also completed. Results: Comparison of ALS and control groups revealed a reduction in CSF flow magnitude and increased flow propagation velocities in the ALS cohort. Other differences in spectral harmonic content and geometric comparisons may support an overall decrease in intrathecal compliance in the ALS group. Notably, there was a high degree of variability between cases, with one ALS patient displaying nearly zero CSF flow along the entire spinal canal. Conclusion: While our sample size limits statistical confidence about the differences observed in this study, it was possible to measure and quantify inter-individual and cohort variability in a non-invasive manner. Our study also shows the potential for MRI based measurements of CSF geometry and flow to provide information about the hydrodynamic environment of the spinal subarachnoid space. These dynamics may be studied further to understand the behavior of CSF solute transport in healthy and diseased states.

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Section: Potential Implications For Intrathecal Als Therapeuticsmentioning

confidence: 99%

Non-invasive MRI quantification of cerebrospinal fluid dynamics in amyotrophic lateral sclerosis patients

Sass

Khani

Romm

et al. 2020

Fluids Barriers CNS

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“… Human artificial chromosome (HAC). ( a ) Diagram of the alphoid tetO -HAC 37 ( tetO -HAC) used in this study. The HAC contains a unique gene-loading loxP site.…”

Section: Figurementioning

confidence: 99%

“… 40 The HAC consists of ~6,000 copies of the 42-bp tetracycline operator (tetO) sequence incorporated into every second alphoid DNA monomer of the 1.1 megabase-size alphoid DNA array. 37 , 43 The HAC has ~30 copies of the selectable marker blasticidin (bsr). ( b ) FISH of a donor hamster CHO cells and recipient human cell lines carrying the autonomously propagated HAC.…”

Section: Figurementioning

confidence: 99%

“…Many publications have described the application of MMCT to transfer MACs/HACs carrying certain genes to gene-deficient cells for gene function studies, humanized animal transgenesis, and high-yield protein production. 21–25 , 28 , 32–37 As MMCT has numerous applications, a more efficient procedure for chromosomes or MACs/HACs transfer is a worthy goal.…”

Section: Introductionmentioning

confidence: 99%

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Moving toward a higher efficiency of microcell-mediated chromosome transfer

Liskovykh

Lee

Larionov

et al. 2016

Molecular Therapy - Methods & Clinical Development

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Microcell-mediated chromosome transfer (MMCT) technology enables individual mammalian chromosomes, megabase-sized chromosome fragments, or mammalian artificial chromosomes that include human artificial chromosomes (HACs) and mouse artificial chromosomes (MACs) to be transferred from donor to recipient cells. In the past few decades, MMCT has been applied to various studies, including mapping the genes, analysis of chromosome status such as aneuploidy and epigenetics. Recently, MMCT was applied to transfer MACs/HACs carrying entire chromosomal copies of genes for genes function studies and has potential for regenerative medicine. However, a safe and efficient MMCT technique remains an important challenge. The original MMCT protocol includes treatment of donor cells by Colcemid to induce micronucleation, where each chromosome becomes surrounded with a nuclear membrane, followed by disarrangement of the actin cytoskeleton using Cytochalasin B to help induce microcells formation. In this study, we modified the protocol and demonstrated that replacing Colcemid and Cytochalasin B with TN-16 + Griseofulvin and Latrunculin B in combination with a Collage/Laminin surface coating increases the efficiency of HAC transfer to recipient cells by almost sixfold and is possibly less damaging to HAC than the standard MMCT method. We tested the improved MMCT protocol on four recipient cell lines, including human mesenchymal stem cells and mouse embryonic stem cells that could facilitate the cell engineering by HACs.

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“…4 Taking advantage of these features, MSC have been employed as tumoraccumulating cells for anticancer therapy in various mouse models. [5][6][7][8][9] Although several studies have combined human MSC and immunodeficient mice, few studies have developed models with mouse MSC and syngeneic mouse tumors. Syngeneic tumor models are critical for investigating in vivo antitumor T cell immunity after MSC therapy.…”

Section: Introductionmentioning

confidence: 99%

Local injection of CCL19-expressing mesenchymal stem cells augments the therapeutic efficacy of anti-PD-L1 antibody by promoting infiltration of immune cells

Iida

Yoshikawa

Murata

et al. 2020

J Immunother Cancer

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BackgroundMesenchymal stem/stromal cells (MSC) accumulate and reside in tumor sites.MethodsTaking advantage of this feature in anticancer therapy, immortalized murine MSC (iMSC) were genetically altered to produce chemokine (C-C motif) ligand 19 (iMSC/CCL19), which attracts dendritic cells (DC) and T lymphocytes. Thereafter, iMSC/CCL19 were examined for their therapeutic efficacy using a syngeneic CT26 colon carcinoma cell line.ResultsCo-injection of iMSC/CCL19 into mice significantly suppressed the in vivo growth of CT26 cells compared with that of CCL19-expressing immortalized fibroblasts (iFib/CCL19). This anticancer effect was not observed when injected in CT26-bearing nude mice. Co-injected iMSC/CCL19 survived longer than iFib/CCL19 in the tumor sites. In a therapeutic model, local injection of iMSC/CCL19 suppressed the tumor growth, and increased IFN (interferon)-γ+ CD8+ T cells and CCR7+ DC infiltration in tumor site was observed when treated with iMSC/CCL19, but not with iMSC. This antitumor effect was completely negated by depletion of CD4+ cells and partially negated by depletion of CD8+ cells. Furthermore, the antitumor effects induced by local injection of iMSC/CCL19 were augmented by additional therapy with anti-programmed death (PD)-ligand 1 (PD-L1) antibody, but not with anti-PD-1 antibody. This combination therapy cured most of the tumors in CT26-bearing mice.ConclusionThese results suggest that local therapy with iMSC/CCL19 can suppress tumor growth via effective recruitment of CCR7+ DC into tumor sites and increase IFN-γ+ CD8+ T cells, and that combination with anti-PD-L1 antibody therapy can be a powerful anticancer therapy.

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Use of a Human Artificial Chromosome for Delivering Trophic Factors in a Rodent Model of Amyotrophic Lateral Sclerosis

Cited by 20 publications

References 22 publications

Non-invasive MRI quantification of cerebrospinal fluid dynamics in amyotrophic lateral sclerosis patients

Non-invasive MRI quantification of cerebrospinal fluid dynamics in amyotrophic lateral sclerosis patients

Moving toward a higher efficiency of microcell-mediated chromosome transfer

Local injection of CCL19-expressing mesenchymal stem cells augments the therapeutic efficacy of anti-PD-L1 antibody by promoting infiltration of immune cells

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