Top

Pediatric Surgery International

Published in:

01-12-2023 | Original Article

The impact of the recipient intestinal site on the differentiation of transplanted enteric neural crest cells

Authors: Nana Nakazawa-Tanaka, Naho Fujiwara, Katsumi Miyahara, Chihiro Akazawa, Masahiko Urao, Atsuyuki Yamataka

Published in: Pediatric Surgery International | Issue 1/2023

Abstract

Purpose

It has long been established that the failure of enteric neural crest cells (ENCCs) to colonize the entire gut results in aganglionosis at the distal colon in Hirschsprung disease (HD). However, it is still unclear how the intestinal microenvironment of the distal aganglionic gut differs from that of the proximal ganglionic gut in HD versus normal gut. We have recently succeeded in transplanting ENCC into aganglionic gut in endothelin receptor B (Ednrb) knockout (KO) mice. to advance the development of cell therapy for HD, it is essential to determine if the transplanted ENCCs differentiate normally in aganglionic gut. Therefore, we designed this study to investigate the impact of the environment of the recipient intestinal tract, at various sites of aganglionic gut, on the differentiation of transplanted ENCCs.

Methods

ENCCs were isolated from Sox10 Venus transgenic (Tg) mouse gut on embryonic day 18.5 (E18.5) and neurospheres (NS) were generated. Then, NS were transplanted into aganglionic KO and wildtype (WT) gut that had been transected just distal to the ENCC wavefront (KO-wf: n = 6, WT: n = 7), and into distal KO gut transected at a site equivalent to that of the WT (KO-d: n = 6) on E12.5. ENCC differentiation was evaluated using whole-mount immunohistochemistry with Tuj-1 (neuronal marker) and GFAP (glial marker) antibodies.

Results

The transplanted ENCCs migrated to form the myenteric and submucosal plexus in all groups. The ratio of the area of Tuj-1-positive cells/GFAP-positive cells in migrated cells in the recipient gut was found to be significantly lower in KO-d compared to KO-wf and WT, while there was no significant difference between KO-wf and WT groups. This suggests that neuronal/glial differentiation was decreased in KO-d compared to that in KO-wf and WT groups.

Conclusion

Our study highlights the differences in ENCC differentiation depending on the site of transplantation. To further develop cell therapy for HD, it is important to consider the impact of the recipient intestinal environment on transplanted ENCCs.

Hei Ha JL, Hang Lui VC, Tam H et al (2022) Embryology and anatomy of Hirschsprung disease PK. Semin Pediatr Surg. https://doi.org/10.1016/j.sempedsurg.2022.151227CrossRefPubMed

Young HM, Bergner AJ, Anderson RB et al (2004) Dynamics of neural crest-derived cell migration in the embryonic mouse gut. Dev Biol 270:455–473CrossRefPubMed

Nakazawa-Tanaka N, Fujiwara N, Miyahara K et al (2022) Increased enteric neural crest cell differentiation after transplantation into aganglionic mouse gut. Pediatr Surg Int. https://doi.org/10.1007/s00383-022-05324-7CrossRefPubMed

Shibata S, Yasuda A, Renault-Mihara F et al (2010) Sox10-Venus mice: a new tool for real-time labeling of neural crest lineage cells and oligodendrocytes. Mol Brain 3:31CrossRefPubMedPubMedCentral

Alhawaj AF (2021) Stem cell-based therapy for hirschsprung disease, do we have the guts to treat? Gene Ther 29:578–587CrossRefPubMedPubMedCentral

Pawolski V, Schmidt MHH (2021) Neuron–Glia interaction in the developing and adult enteric nervous system. Cells 10(1):47. https://doi.org/10.3390/cells10010047CrossRef

Liu JA, Ngan ES (2014) Hedgehog and Notch signaling in enteric nervous system development. Neurosignals 22:1–13. https://doi.org/10.1159/000356305CrossRefPubMed

Young HM, Bergner AJ, Muller T (2003) Acquisition of neuronal and glial markers by neural crest-derived cells in the mouse intestine. J Comp Neurol 456:1–11CrossRefPubMed

Boesmans W, Nash A, Tasnády KR et al (2022) Development, diversity, and neurogenic capacity of enteric glia. Front Cell Dev Biol 17(9):775102. https://doi.org/10.3389/fcell.2021.775102CrossRef

10.

Soret R, Schneider S, Bernas G et al (2022) Glial cell-derived neurotrophic factor induces enteric neurogenesis and improves colon structure and function in mouse models of hirschsprung disease. Gastroenterology 159:1824–1838. https://doi.org/10.1053/j.gastro.2020.07.018CrossRef

11.

Graham KD, López SH, Sengupta R et al (2020) Robust, 3-dimensional visualization of human colon enteric nervous system without tissue sectioning. Gastroenterology 158:2221–2235. https://doi.org/10.1053/j.gastro.2020.02.035CrossRefPubMed

12.

Badizadegan K, Thomas AR, Nagy N et al (2014) Presence of intramucosal neuroglial cells in normal and ganglionic human colon. Am J Physiol Gastrointest Liver Physiol 15(307):G1002–G1012. https://doi.org/10.1152/ajpgi.00164.2014CrossRef

13.

Veríssimo CP, Carvalho JDS, da Silva FJM et al (2019) Laminin and environmental cues act in the inhibition of the neuronal differentiation of enteric glia in vitro. Front Neurosci 3:914. https://doi.org/10.3389/fnins.2019.00914CrossRef

14.

Payette RF, Tennyson VM, Pomeranz HD et al (1988) Accumulation of components of basal laminae: association with the failure of neural crest cells to colonize the presumptive aganglionic bowel of ls/ls mutant mice. Dev Biol 125:341–360CrossRefPubMed

15.

Parikh DH, Tam PK, Lloyd DA et al (1992) Quantitative and qualitative analysis of the extracellular matrix protein, laminin, in Hirschsprung’s disease. J Pediatr Surg 27:991–996CrossRefPubMed

16.

Li AW, Zhang WT, Wang R et al (2008) The relationship between expressions of the laminin gene and RET gene in Hirschsprung’s disease. World J Pediatr 4:135–139CrossRefPubMed

17.

Fujiwara N, Nakazawa-Tanaka N, Miyahara K et al (2017) Altered expression of laminin alpha1 in aganglionic colon of endothelin receptor-B null mouse model of Hirschsprung’s disease. Pediatr Surg Int 34:137–141. https://doi.org/10.1007/s00383-017-4180-6CrossRefPubMed

Title: The impact of the recipient intestinal site on the differentiation of transplanted enteric neural crest cells
Authors: Nana Nakazawa-Tanaka
Naho Fujiwara
Katsumi Miyahara
Chihiro Akazawa
Masahiko Urao
Atsuyuki Yamataka
Publication date: 01-12-2023
Publisher: Springer Berlin Heidelberg
Published in: Pediatric Surgery International / Issue 1/2023
Print ISSN: 0179-0358
Electronic ISSN: 1437-9813
DOI: https://doi.org/10.1007/s00383-023-05587-8

At a glance: The STEP trials

Springer Medicine

The impact of the recipient intestinal site on the differentiation of transplanted enteric neural crest cells

Abstract

Purpose

Methods

Results

Conclusion

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2023

Preexisting neural factors that contribute to dysmotility in esophageal atresia: a systematic review

Anastomotic stricture prediction in patients with esophageal atresia with distal fistula

A nomogram for the preoperative estimation of neuroblastoma risk despite inadequate biopsy information

Stump appendicitis in children: a systematic review

Parental perceptions and experiences of care in the surgical neonatal intensive care unit

Long-term bowel function after single-stage transanal endorectal pull-through in neonatal patients with Hirschsprung disease