Tuft Cells: A New Player in Hirschsprung's Disease

Abstract

Introduction “Tuft” cells, also known as brush or caveolated cells, are characteristically fusiform shaped, with a distinct apical “tuft” of microvilli extending into the lumen. Double cortin-like kinase 1 (DCLK1) is a microtubule kinase and is a specific marker of intestinal tuft cells. DCLK1-positive tuft cells have been shown to play a key role in gastrointestinal chemosensation, inflammation, and neurotransmission. DCLK1 and Choline acetyltransferase (ChAT), the enzymes responsible for acetylcholine production, are reported to be coexpressed within the gastrointestinal tract. We designed this study to investigate the hypothesis that DCLK1 gene expression is altered in Hirschsprung's disease (HSCR).

Materials and Methods HSCR tissue specimens (n = 6) were collected at the time of pull-through surgery, while control samples were obtained at the time of colostomy closure in patients with imperforate anus (n = 6). Quantitative real-time polymerase chain reaction (qRT-PCR) analysis was undertaken to quantify DCLK1 gene expression, and immunolabeling of DCLK1-positive tuft cells was visualized using confocal microscopy.

Results qRT-PCR analysis revealed significant downregulation of the DCLK1 gene in both aganglionic and ganglionic HSCR specimens compared with controls (p < 0.05). Confocal microscopy revealed DCLK1-positive tuft cell expression within the colonic mucosa, with a reduction in expression in both aganglionic and ganglionic HSCR colon compared with controls.

Conclusion DCLK1 is significantly downregulated in HSCR colon, suggesting a role for tuft cells in cholinergic neurotransmission of the distal colon. The marked decrease in DCLK1 expression within ganglionic specimens highlights the physiologically abnormal nature of this segment in HSCR patients.

Keywords

tuft cells - double cortin-like kinase 1 - Hirschsprung's disease - aganglionosis

• References

• 1 Schütz B, Jurastow I, Bader S. , et al. Chemical coding and chemosensory properties of cholinergic brush cells in the mouse gastrointestinal and biliary tract. Front Physiol 2015; 6: 87
• 2 Yi J, Bergstrom K, Fu J. , et al. Dclk1 in tuft cells promotes inflammation-driven epithelial restitution and mitigates chronic colitis. Cell Death Differ 2019; 26 (09) 1656-1669
• 3 Leppänen J, Helminen O, Huhta H. , et al. Doublecortin-like kinase 1-positive enterocyte - a new cell type in human intestine. APMIS 2016; 124 (11) 958-965
• 4 Menezes M, Corbally M, Puri P. Long-term results of bowel function after treatment for Hirschsprung's disease: a 29-year review. Pediatr Surg Int 2006; 22 (12) 987-990
• 5 Menezes M, Pini Prato A, Jasonni V, Puri P. Long-term clinical outcome in patients with total colonic aganglionosis: a 31-year review. J Pediatr Surg 2008; 43 (09) 1696-1699
• 6 Laughlin DM, Friedmacher F, Puri P. Total colonic aganglionosis: a systematic review and meta-analysis of long-term clinical outcome. Pediatr Surg Int 2012; 28 (08) 773-779
• 7 O'Donnell AM, Coyle D, Puri P. Decreased expression of hyperpolarisation-activated cyclic nucleotide-gated channel 3 in Hirschsprung's disease. World J Gastroenterol 2015; 21 (18) 5635-5640
• 8 O'Donnell AM, Coyle D, Puri P. Deficiency of platelet-derived growth factor receptor-α-positive cells in Hirschsprung's disease colon. World J Gastroenterol 2016; 22 (12) 3335-3340
• 9 O'Donnell AM, Nakamura H, Tomuschat C, Marayati NF, Puri P. Altered expression of KCNG3 and KCNG4 in Hirschsprung's disease. Pediatr Surg Int 2019; 35 (02) 193-197
• 10 O'Donnell AM, Nakamura H, Tomuschat C, Marayati NF, Puri P. Abnormal Scn1b and Fxyd1 gene expression in the pulled-through ganglionic colon may influence functional outcome in patients with Hirschsprung's disease. Pediatr Surg Int 2019; 35 (01) 9-14
• 11 Cheng X, Voss U, Ekblad E. Tuft cells: distribution and connections with nerves and endocrine cells in mouse intestine. Exp Cell Res 2018; 369 (01) 105-111
• 12 Middelhoff M, Westphalen CB, Hayakawa Y. , et al. Dclk1-expressing tuft cells: critical modulators of the intestinal niche?. Am J Physiol Gastrointest Liver Physiol 2017; 313 (04) G285-G299
• 13 Cheng X, Voss U, Ekblad E. A novel serotonin-containing tuft cell subpopulation in mouse intestine. Cell Tissue Res 2019; 376 (02) 189-197
• 14 Westphalen CB, Asfaha S, Hayakawa Y. , et al. Long-lived intestinal tuft cells serve as colon cancer-initiating cells. J Clin Invest 2014; 124 (03) 1283-1295