Optimal route of diphtheria toxin administration to eliminate native nephron progenitor cells in vivo for kidney regeneration

Fukunaga, Shohei; Yamanaka, Shuichiro; Fujimoto, Toshinari; Tajiri, Susumu; Uchiyama, Taketo; Matsumoto, Keīichi; Ito, Takafumi; Tanabe, Kazuaki; Yokoo, Takashi

doi:10.1016/j.bbrc.2018.01.166

Cited by 7 publications

(3 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To address that, a low-nephron number model using the Six2/iDTR model has been proposed by the author ( 61 ). This model expresses the diphtheria toxin receptor in nephron progenitor cells, which can be removed by administering diphtheria toxin to the amniotic fluid during the embryonic period.…”

Section: Interventions To Preserve Nephron Number At Birthmentioning

confidence: 99%

Low glomerular number at birth can lead to the development of chronic kidney disease

Fukunaga

Fujita

2023

Front. Endocrinol.

Self Cite

View full text Add to dashboard Cite

Chronic kidney disease (CKD) prevalence is increasing worldwide, and reducing the number of patients with CKD is of utmost importance. The environment during the fetal, perinatal, and early childhood stages may influence CKD development (developmental origins of health and disease). Under conditions of maternal malnutrition, the glomerular number of infants reduces, and the risk of developing CKD may increase. Nephron progenitor cells and ureteric buds interact with each other to form glomeruli at the tip of the ureteric bud. Thus, the number of glomeruli is determined by the number of ureteric bud branches, which are reportedly decreased due to maternal malnutrition, in turn reducing the glomerular number. Four possible mechanisms can explain the low glomerular number resulting from maternal malnutrition: 1) suppression of c-Ret expression, 2) suppression of nephron formation by renin-angiotensin-aldosterone system inhibition, 3) exposure to excess glucocorticoids, and 4) promotion of apoptosis. Additionally, nephron formation does not continue after birth in humans. Therefore, a low glomerular number at birth is a lifelong burden on the glomeruli and increases the risk of developing CKD. Therefore, it is important to maintain the glomerular number at birth. Accurate glomerular counts are essential for conducting studies on the glomerular number. The dissector/fractionator method is the gold standard; however, it can only be performed at some institutions. Recently, methods have been developed to measure the glomerular number by combining computed tomography and pathological examination and measure the glomerular count using magnetic resonance imaging. Models of decreased and increased glomerular numbers have been developed. Moreover, research regarding the causes of decreased glomerular number and its relationship with development of lifestyle-related diseases and renal dysfunction has significantly progressed, furthering our understanding of the importance of glomerular number.

show abstract

Section: Interventions To Preserve Nephron Number At Birthmentioning

confidence: 99%

Low glomerular number at birth can lead to the development of chronic kidney disease

Fukunaga

Fujita

2023

Front. Endocrinol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…intraperitoneal) or local (e.g. subcutaneous, intratracheal) administration (17)(18)(19)(20). Using three different DT doses in AAV-stuffer control animals we measured a DT dose dependent influx of neutrophils and macrophages and correspondingly also increased concentrations of MIPs.…”

Section: Dt Off-target Effectsmentioning

confidence: 99%

Characterization of a flexible AAV-DTR/DT mouse model of acute epithelial lung injury

Griesser

Schönberger

Stierstorfer

et al. 2022

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

Animal models are important to mimic certain pathways or biological aspects of human pathologies including acute and chronic pulmonary diseases. We developed a novel and flexible mouse model of acute epithelial lung injury based on adeno-associated virus (AAV) variant 6.2 mediated expression of the human diphtheria toxin receptor (DTR). Following intratracheal administration of diphtheria toxin (DT), a cell-specific death of bronchial and alveolar epithelial cells can be observed. In contrast to other lung injury models, the here described mouse model provides the possibility of targeted injury using specific tropisms of AAV vectors or cell type specific promotors to drive the human DTR expression. Also, generation of cell specific mouse lines is not required. Detailed characterization of the AAV-DTR/DT mouse model including titration of viral genome (vg) load and administered DT amount revealed increasing cell numbers in bronchoalveolar lavage (BAL; macrophages, neutrophils, and unspecified cells) and elevation of degenerated cells and infiltrated leukocytes in lung tissue, dependent of vg load and DT dose. Cytokine levels in BAL fluid showed different patterns with higher vg load, e.g. IFNγ, TNFα, and IP10 increasing and IL-5 and IL-6 decreasing, while lung function was not affected. Additionally, laser-capture microdissection (LCM)-based proteomics of bronchial epithelium and alveolar tissue revealed upregulated immune and inflammatory response in all regions and extracellular matrix deposition in infiltrated alveoli. Overall, our novel AAV-DTR/DT model allows investigation of repair mechanisms following epithelial injury and resembles specific mechanistic aspects of acute and chronic pulmonary diseases.

show abstract

“…Details for this procedure have been described previously. 22 A diphtheria toxin (DT) (50 ng/ml; Wako) stock solution, stored at 280°C until use, was thawed and diluted in PBS. Pregnant female mice were anesthetized and laparotomized; in each case, the uterus was delivered through the incision.…”

Section: Diphtheria Toxin Injection Into Amniotic Fluidmentioning

confidence: 99%

Kidney Regeneration in Later-Stage Mouse Embryos via Transplanted Renal Progenitor Cells

Yamanaka

Saito

Fujimoto

et al. 2019

JASN

Self Cite

View full text Add to dashboard Cite

BackgroundThe limited availability of donor kidneys for transplantation has spurred interest in investigating alternative strategies, such as regenerating organs from stem cells transplanted into animal embryos. However, there is no known method for transplanting cells into later-stage embryos, which may be the most suitable host stages for organogenesis, particularly into regions useful for kidney regeneration.MethodsWe demonstrated accurate transplantation of renal progenitor cells expressing green fluorescent protein to the fetal kidney development area by incising the opaque uterine muscle layer but not the transparent amniotic membrane. We allowed renal progenitor cell–transplanted fetuses to develop for 6 days postoperatively before removal for analysis. We also transplanted renal progenitor cells into conditional kidney-deficient mouse embryos. We determined growth and differentiation of transplanted cells in all cases.ResultsRenal progenitor cell transplantation into the retroperitoneal cavity of fetuses at E13–E14 produced transplant-derived, vascularized glomeruli with filtration function and did not affect fetal growth or survival. Cells transplanted to the nephrogenic zone produced a chimera in the cap mesenchyme of donor and host nephron progenitor cells. Renal progenitor cells transplanted to conditional kidney-deficient fetuses induced the formation of a new nephron in the fetus that is connected to the host ureteric bud.ConclusionsWe developed a cell transplantation method for midstage to late-stage fetuses. In vivo kidney regeneration from renal progenitor cells using the renal developmental environment of the fetus shows promise. Our findings suggest that fetal transplantation methods may contribute to organ regeneration and developmental research.

show abstract

Optimal route of diphtheria toxin administration to eliminate native nephron progenitor cells in vivo for kidney regeneration

Cited by 7 publications

References 25 publications

Low glomerular number at birth can lead to the development of chronic kidney disease

Low glomerular number at birth can lead to the development of chronic kidney disease

Characterization of a flexible AAV-DTR/DT mouse model of acute epithelial lung injury

Kidney Regeneration in Later-Stage Mouse Embryos via Transplanted Renal Progenitor Cells

Contact Info

Product

Resources

About