Epithelial Regeneration Ability of Crohn's Disease Assessed Using Patient-Derived Intestinal Organoids

Abstract Background: Little is known about the ability of epithelial regeneration and wound healing in patients with inflammatory bowel disease, since the ethical issue of clinical trials and the inherent limitations of tumor-derived epithelial cell lines and animal models. We evaluated the epithelial proliferation and wound healing ability of the patients with Crohn’s disease (CD) using the patient-derived intestinal organoids. Methods: Human intestinal organoids were constructed in a three-dimensional intestinal crypt culture of enteroscopic biopsy samples from controls and CD patients. The epithelial regeneration ability of intestinal organoids was assessed using organoid reconstitution, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), 5-ethynyl-2′-deoxyuridine (EdU), and wound healing assays. Tumor necrosis factor-alpha (TNFa) is a major pro-inflammatory effector during the CD pathogenesis. Under the TNFa-enriched conditions, the epithelial regeneration ability of CD patient-derived organoids was compared with that of control organoids.Results: The organoid-forming efficiency of ileal crypts derived from CD patients was reduced compared with those from control subjects (p < .001). Long-term cultured organoids (≥ 6 passages) derived from controls and CD patients showed inseparable morphologic and cultural features. Under the TNFa-enriched conditions, organoid reconstitution assay and MTT assay revealed that the organoid reconstitution and viability of CD patient-derived organoids were significantly lower than those of control organoids (p <.05 for each). The number of EdU+ proliferative cells was significantly lower in TNFα-treated organoids derived from CD patients than in TNFα-treated control organoids (p < .05). In the wound healing assay, the unhealed area in TNFα-treated CD patient-derived organoids was significantly larger than that of TNFα-treated control organoids (p < .001). Conclusions: Under TNFα-enriched conditions, the wound healing ability of CD patient-derived organoids is reduced due to the reduced cell proliferation compared with that of control organoids. The epithelial regeneration ability may be impaired in patients with CD.

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Epithelial Regeneration Ability of Crohn’s Disease Assessed Using Patient-Derived Intestinal Organoids

International Journal of Molecular Sciences ◽

10.3390/ijms22116013 ◽

2021 ◽

Vol 22 (11) ◽

pp. 6013

Author(s):

Chansu Lee ◽

Sung-Noh Hong ◽

Eun-Ran Kim ◽

Dong-Kyung Chang ◽

Young-Ho Kim

Keyword(s):

Wound Healing ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Three Dimensional ◽

Regeneration Ability ◽

Microscopic Appearance ◽

Epithelial Regeneration ◽

Intestinal Organoids ◽

Bowel Diseases ◽

Forming Efficiency

Little is known about the ability for epithelial regeneration and wound healing in patients with inflammatory bowel diseases. We evaluated the epithelial proliferation and wound healing ability of patients with Crohn’s disease (CD) using patient-derived intestinal organoids. Human intestinal organoids were constructed in a three-dimensional intestinal crypt culture of enteroscopic biopsy samples from controls and CD patients. The organoid-forming efficiency of ileal crypts derived from CD patients was reduced compared with those from control subjects (p < 0.001). Long-term cultured organoids (≥6 passages) derived from controls and CD patients showed an indistinguishable microscopic appearance and culturing behavior. Under TNFα-enriched conditions (30 ng/mL), the organoid reconstitution rate and cell viability of CD patient-derived organoids were significantly lower than those of the control organoids (p < 0.05 for each). The number of EdU+ cells was significantly lower in TNFα-treated organoids derived from CD patients than in TNFα-treated control organoids (p < 0.05). In a wound healing assay, the unhealed area in TNFα-treated CD patient-derived organoids was significantly larger than that of TNFα-treated control organoids (p < 0.001). The wound healing ability of CD patient-derived organoids is reduced in TNFα-enriched conditions, due to reduced cell proliferation. Epithelial regeneration ability may be impaired in patients with CD.

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Epithelial Regeneration Ability of Crohn's Disease Assessed Using Patient-Derived Intestinal Organoids

10.1101/2021.03.25.437111 ◽

2021 ◽

Author(s):

Chnasu Lee ◽

Sung Noh Hong ◽

Eun Ran Kim ◽

Dong Kyung Chang ◽

Young-Ho Kim

Keyword(s):

Wound Healing ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Wound Repair ◽

Ex Vivo ◽

Formation Rate ◽

Regeneration Ability ◽

Epithelial Regeneration ◽

Control Subjects ◽

Intestinal Organoids

Background: There are limitations to the utility of tumor-derived epithelial cell lines and animal models to evaluate the proliferation ability and wound repair of intestinal epithelium in the context of Crohn's disease (CD). Therefore, we aimed to study epithelial regeneration ability in CD using an intestinal organoid model. Further, since tumor necrosis factor alpha (TNFα) is a major proinflammatory effector during CD pathogenesis, we also investigated TNFα-induced alteration of regeneration ability in CD patient-derived intestinal organoids. Methods: Human intestinal organoids were constructed in a three-dimensional intestinal crypt culture of enteroscopic biopsy samples from control subjects and patients with CD. The epithelial regeneration ability of intestinal organoids was assessed using organoid reconstitution, 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), and wound healing assays. Results: Ex vivo cultures of ileal crypt cells revealed that organoid formation rate of CD patients were reduced compared with that of control subjects (p <.001). CD patient-derived organoids sub-cultured for more than 6 passages showed stable organoid reconstitution and identical morphological features. The organoid constitution and MTT assay revealed that the viability of TNFα-treated CD patient-derived organoids were significantly lower than that of TNFα-treated control organoids (p <.05 for each). The number of EdU+ proliferative cells was significantly lower in TNFα-treated CD patient-derived organoids than in TNFα-treated control organoids (p <.05). The wound-healing ability of TNFα-treated CD patient-derived organoids was significantly lower than that of TNFα-treated control organoids (p <.001). Conclusions: The ability of proliferation and wound healing in CD-patients derived organoids is reduced, especially in TNFα-enriched condition. Our results indicated that the epithelial regenerative ability may be impaired in patients with CD under inflammatory conditions.

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Infliximab: A Review of its use in the Treatment of Pediatric Crohn's Disease

Clinical Medicine Insights Therapeutics ◽

10.4137/cmt.s2840 ◽

2010 ◽

Vol 2 ◽

pp. CMT.S2840

Author(s):

Arkady Broder ◽

Joel R. Rosh

Keyword(s):

Monoclonal Antibodies ◽

Crohn’S Disease ◽

Chronic Disease ◽

Crohn's Disease ◽

Tumor Necrosis ◽

Immune Dysregulation ◽

Necrosis Factor Alpha ◽

The Past ◽

Factor Alpha ◽

Necrosis Factor

The past decade has brought great change to the treatment of pediatric Crohn's disease. The majority of affected patients now receive therapy directed at the underlying immune dysregulation that is associated with this chronic disease. The monoclonal antibodies directed against tumor necrosis factor alpha play an increasing role in such therapy. Infliximab is the prototype of this class of biologic based therapy. This review covers the basic pharmacokinetics of infliximab while reviewing the data on its efficacy in pediatric Crohn's disease patients. Current issues related to infliximab dosing and safety are also reviewed.

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Differential Immunohistochemical Expression of Syndecan-1 and Tumor Necrosis Factor Alpha in Colonic Mucosa of Patients with Crohn’s Disease

Immunopharmacology and Immunotoxicology ◽

10.1080/a08923970600815048 ◽

2006 ◽

Vol 28 (2) ◽

pp. 185-195 ◽

Cited By ~ 4

Author(s):

Mariabeatrice Principi ◽

Richard Day ◽

Stefania Marangi ◽

Osvaldo Burattini ◽

Vincenzo De Francesco ◽

...

Keyword(s):

Crohn’S Disease ◽

Tumor Necrosis Factor ◽

Crohn's Disease ◽

Tumor Necrosis Factor Alpha ◽

Tumor Necrosis ◽

Colonic Mucosa ◽

Immunohistochemical Expression ◽

Necrosis Factor Alpha ◽

Factor Alpha ◽

Necrosis Factor

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935 PRE-TREATMENT MUCOSAL INFLAMMATORY AND WOUND HEALING GENE PROGRAMS REVEAL MECHANISMS ASSOCIATED WITH FUTURE STRICTURING BEHAVIOR DURING FIVE YEAR FOLLOW-UP IN PEDIATRIC CROHN'S DISEASE

Gastroenterology ◽

10.1016/s0016-5085(20)31152-5 ◽

2020 ◽

Vol 158 (6) ◽

pp. S-187-S-188

Author(s):

Yael Haberman ◽

Phillip P. Minar ◽

Rebekah Karns ◽

Phillip J. Dexheimer ◽

Sudhir Ghandikota ◽

...

Keyword(s):

Wound Healing ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Pre Treatment

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Tumor necrosis factor-alpha antibody for induction of remission in Crohn's disease

10.1002/14651858.cd003574 ◽

2001 ◽

Cited By ~ 1

Author(s):

AK Akobeng ◽

M Zachos

Keyword(s):

Crohn’S Disease ◽

Tumor Necrosis Factor ◽

Crohn's Disease ◽

Tumor Necrosis Factor Alpha ◽

Tumor Necrosis ◽

Necrosis Factor Alpha ◽

Factor Alpha ◽

Necrosis Factor

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P066 Azathioprine linked with impaired intestinal epithelial regeneration in Crohn’s disease

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz203.195 ◽

2020 ◽

Vol 14 (Supplement_1) ◽

pp. S167-S168

Author(s):

K Borycka-Kiciak ◽

A Pietrzak ◽

K Ferenc ◽

P Pietrzak ◽

L Janaszek ◽

...

Keyword(s):

Dna Damage ◽

Crohn’S Disease ◽

Crohn's Disease ◽

Epithelial Cells ◽

Caspase 3 ◽

Surrogate Marker ◽

Preoperative Treatment ◽

Intestinal Epithelial ◽

Epithelial Regeneration ◽

E Cadherin

Abstract Background In patients operated due to active Crohn’s disease (CD), the negative effect of steroids and biologics on septic complications is known. Until now, azathioprine (AZA) is considered to be safe. The aim of our study was to assess the impact of AZA on intestinal epithelial cells damage in patients with active CD as a surrogate marker of healing. Methods Intestinal specimens taken from macroscopically healthy surgical margins of all consecutive CD patients operated due to active isolated ileocecal disease during the study period (2014–2016) were evaluated. Expression of caspase-3, p-53 and Ki-67 as markers of cell apoptosis, DNA damage and proliferation were immunohistochemically tested and assessed using a confocal microscope and microimage for in-tissue-cytometry analysis. Western-blot analysis was performed for the evaluation of cellular integrity using ZO-1 and E-cadherin as a markers. 30-day clinical outcomes were assessed. The study was approved by the institutional Ethics Committee. Results From 61 operated due to active CD patients, 35 met the inclusion criteria. Patients were divided accordingly to preoperative treatment: treated with no immunomodulators (N-9 patients), on steroids (S-14 patients), on AZA (A-6 patients), and on combination therapy, AZA + steroids (AS-6 patients). There were no substantial differences between groups. We found statistically significant increase of apoptosis in A group compared with N (5.33 ± 1.05 vs. 1.29 ± 0.51, p = 0.011), but also S group (1.58 ± 0.68, p = 0.014) and increase of DNA damage in A, AS and S groups compared with N group (p =0.01; p = 0.032; p = 0.035, respectively) (Figure 1a–c). P53-mediated cell cycle arrest and apoptosis through a caspase-3-dependent pathway in response to DNA damage was the most intensive in A group (Figure 2a). Reduction of cell proliferative activity in group A did not reach statistical significance (p = 0,057). A reduction of ZO-1 in A group and increased level of E-cadherin in S group were found. The effect of the decreased number of tight junctions and disintegration of the mucosa layer was observed in A group (Figure 2b). Clinically, in 30-day postoperative follow-up, six wound healing complications and one anastomotic leak were found, all in patients treated with immunomodulators. Conclusion We found that in epithelial cells of the small and large intestine of patients treated with AZA, apoptotic activity and DNA damage processes are increased when regeneration processes and mucosal integrity are significantly disturbed. These abnormalities of intestinal epithelial regeneration may be a surrogate marker of impaired mucosal healing.

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The Cost-effectiveness of Biological Therapy Cycles in the Management of Crohn’s Disease

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz063 ◽

2019 ◽

Vol 13 (10) ◽

pp. 1323-1333 ◽

Cited By ~ 4

Author(s):

Kristian Bolin ◽

Erik Hertervig ◽

Edouard Louis

Keyword(s):

Crohn’S Disease ◽

Cost Effectiveness ◽

Crohn's Disease ◽

Combination Therapy ◽

Willingness To Pay ◽

Cost Effective ◽

Treatment Intensification ◽

Tree Model ◽

Necrosis Factor Alpha ◽

The Cost

Abstract Objectives To examine the cost-effectiveness of continued treatment for patients with moderate-severe Crohn’s disease in clinical remission, with a combination of anti-tumour necrosis factor alpha [anti-TNFα] [infliximab] and immunomodulator therapy compared with two different withdrawal strategies: [1] withdrawal of the anti-TNFα therapy; and [2] withdrawal of the immunomodulator therapy, respectively. Methods A decision-tree model was constructed mimicking three treatment arms: [1] continued combination therapy with infliximab and immunomodulator; [2] withdrawal of infliximab; or [3] withdrawal of the immunomodulator. Relapses in each arm are managed with treatment intensification and re-institution of the de-escalated drug according to a prespecified algorithm. State-dependent relapse risks, remission probabilities, and quality of life weights were collected from previous published studies. Results Combination therapy was less costly and more efficient than the withdrawal of the immunomodulator, and more costly and more efficient than withdrawal of infliximab. Whether or not combination therapy is cost-effective, compared with the alternatives, depends primarily on current pharmaceutical prices and the willingness-to-pay per additional quality-adjusted life-year [QALY]. Conclusions Combination therapy using a combination of anti-TNFα [infliximab] and an immunomodulator is cost-effective in the treatment of Crohn’s disease compared with treatment cycles in which the immunomodulator is withdrawn. Combination treatment is cost-effective compared with treatment cycles in which infliximab is withdrawn, at prices of infliximab below€192/100 mg, given a willingness-to-pay threshold at€49 020 [Sweden] per additional QALY.

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