Factors associated with anti-tumor necrosis factor effectiveness to prevent postoperative recurrence in Crohn’s disease

Anthony Buisson,Lisa Cannon,Konstantin Umanskiy,Roger D. Hurst,Neil H. Hyman,Atsushi Sakuraba,Joel Pekow,Sushila Dalal,Russell D. Cohen,Bruno Pereira,David T. Rubin 대한장연구학회 2022 Intestinal Research Vol.20 No.3

Background/Aims: We assessed the effectiveness of anti-TNF agents and its associated factors to prevent endoscopic and clinical postoperative recurrence (POR) in Crohn’s disease (CD). Methods: From a prospectively-maintained database, we retrieved 316 CD patients who underwent intestinal resection (2011–2017). Endoscopic (Rutgeerts index ≥ i2 at 6 months) and clinical (recurrence of symptoms leading to hospitalization or therapeutic escalation) POR were assessed. Results: In 117 anti-TNF-naïve patients, anti-TNF therapy was more effective than immunosuppressive agents (odds ratio [OR], 8.8; 95% confidence interval [CI], 1.8–43.9; <i>P</i>= 0.008) and no medication/5-aminosalicylates (OR, 5.2; 95% CI, 1.0–27.9; <i>P</i>= 0.05) to prevent endoscopic POR. In 199 patients exposed to anti-TNF prior to the surgery, combination with anti-TNF and immunosuppressive agents was more effective than anti-TNF monotherapy (OR, 2.32; 95% CI, 1.02–5.31; <i>P</i>= 0.046) to prevent endoscopic POR. Primary failure to anti-TNF agent prior to surgery was predictive of anti-TNF failure to prevent endoscopic POR (OR, 2.41; 95% CI, 1.10–5.32; <i>P</i>= 0.03). When endoscopic POR despite anti-TNF prophylactic medication (n = 55), optimizing anti-TNF and adding an immunosuppressive drug was the most effective option to prevent clinical POR (hazard ratio, 7.38; 95% CI, 1.54–35.30; <i>P</i>= 0.012). Anti-TNF therapy was the best option to prevent clinical POR (hazard ratio, 3.10; 95% CI, 1.09–8.83; <i>P</i>= 0.034) in patients with endoscopic POR who did not receive any biologic to prevent endoscopic POR (n = 55). Conclusions: Anti-TNF was the most effective medication to prevent endoscopic and clinical POR. Combination with anti-TNF and immunosuppressive agents should be considered in patients previously exposed to anti-TNF.

Formation of Functional Hepatocyte-Like Cells through Direct Conversion and Transplantation into Various Mice Models

( Sangtae Yoon ),( Kyojin Kang ),( Yohan Kim ),( Elina Maria Buisson ),( Chang Hee Lee ),( Ji-hye Yim ),( Jaemin Jeong ),( Dongho Choi ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: The incidence of liver disease is increasing worldwide. Liver transplantation is the only way to treat serious liver diseases. However, demand for transplants is increasing, while supply is very scarce. Therefore, many researchers are studying ways to replace liver transplantation. The most representative of these is cell therapy using ESCs and iPSCs. However, these are not available for clinical use because of the disadvantage that it may form teratoma in the body. Therefore we suggest other method that direct conversion technique produce generation of hepatocyte-like cells from fibroblasts with two liver-specific transcription factors. Methods: First generation of hepatocyte-like cells (miHeps) were produced by lentivirus infection for continuously expression in host genome during conversion. Second generation of hepatocyte-like cells (R-iHeps) were produced by mRNA transfection. NSG and Albumin-Treck mice were used for in vivo transplantation. Results: Directly converted hepatocyte-like cells can proliferate, split, store, re-seed, and mature with DMSO. And hepatocyte marker genes and protein expressions, albumin, AFP, etc., were increased in both cells, and albumin secretion in the media was much higher than fibroblasts. Also these cells were transplanted into liver injury model, jo2-treated NSG mice and Albumin-Treck mice, to confirm engraftment in liver. After transplant, miHeps and R-Heps can be detected in the liver under the fluorescence microscope. Conclusions: Directly converted hepatocyte-like cells can be useful for liver regeneration instead of ESCs and iPSCs-derived hepatocyte-like cells.

Reprogramming of Human Hepatocytes into Bi-Potent Hepatic Progenitor Cells Using Cocktail of Small Mole-cules and Growth Factor

( Yohan Kim ),( Kyojin Kang ),( Sangtae Yoon ),( Elina Maria Buisson ),( Chang Hee Lee ),( Ji-hye Yim ),( Jaemin Jeong ),( Dongho Choi ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Cell-based regenerative medicine can provide valuable options for end-stage patients with liver disease by solving and considering the lack of donor shortage through gene and/or stem cell therapy. Despite much progress in isolation and prolonged growth of bipotent progenitor cells with regenerative capacity from terminally differentiated mouse hepatocytes, expansion of the adult human hepatocytes remains a major challenge. Methods: We report using two small molecules and growth factors to successfully produce patient-specific hepatic progenitor cells from healthy and diseased human hepatocytes. Results: After three days of treatment small molecule in the presence of growth factor, a key driver of hepatic progenitor cell activity, triggered expansion of small polygonal cells, which co-expressed known hepatic progenitor cells and lineage specific marker genes. These chemically derived human hepatic progenitor cells (hCdHs) could self-renew for at least 10 passages while retaining phenotype, normal karyotype and potential to differentiate into functional hepatocytes and biliary epithelial cells in vitro. A next-generation sequencing confirmed a high degree of molecular similarity between hCdHs and human hepatoblasts. Upon intrasplenic transplantation into immunocompromised mice with a diseased liver, hCdHs effectively repopulated and restored. Conclusions: In conclusion, hCdHs provide a safe novel tool that permits expansion and genetic manipulation of patient-specific hepatic progenitor cells to study regeneration and repair of diseased liver.

Nonintegrating Direct Conversion Using mRNA into Hepatocyte-Like Cells

Yoon, Sangtae,Kang, Kyojin,Cho, Young-duck,Kim, Yohan,Buisson, Elina Maria,Yim, Ji-Hye,Lee, Seung Bum,Ryu, Ki-Young,Jeong, Jaemin,Choi, Dongho Hindawi 2018 BioMed research international Vol.2018 No.-

<P>Recently, several researchers have reported that direct reprogramming techniques can be used to differentiate fibroblasts into hepatocyte-like cells without a pluripotent intermediate step. However, the use of viral vectors for conversion continues to pose important challenges in terms of genome integration. Herein, we propose a new method of direct conversion without genome integration with potential clinical applications. To generate hepatocyte-like cells, mRNA coding for the hepatic transcription factors Foxa3 and HNF4<I>α</I> was transfected into mouse embryonic fibroblasts. After 10-12 days, the fibroblasts converted to an epithelial morphology and generated colonies of hepatocyte-like cells (R-iHeps). The generated R-iHeps expressed hepatocyte-specific marker genes and proteins, including albumin, alpha-fetoprotein, HNF4<I>α</I>, CK18, and CYP1A2. To evaluate hepatic function, indocyanine green uptake, periodic acid-Schiff staining, and albumin secretion were assessed. Furthermore, mCherry-positive R-iHeps were engrafted in the liver of Alb-TRECK/SCID mice, and we confirmed FAH enzyme expression in Fah<SUP>1R</SUP>Tyr<SUP>c</SUP>/RJ models. In conclusion, our data suggest that the nonintegrating method using mRNA has potential for cell therapy.</P>

Chemical Derived Patient-Specific Hepatic Stem Cells from Various Patients

( Kyojin Kang ),( Yohan Kim ),( Sangtae Yoon ),( Elina Maria Buisson ),( Changhee Lee ),( Ji-hye Yim ),( Jaemin Jeong ),( Dongho Choi ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Hepatocyte like cells can be derived from many different sources of stem cells including induced pluripotent stem cells, embryonic stem cells or directly converted cells. These Methods: are well developed, however, some problems such as tumor formation, low functionality and differentiation ratio as well as genomic variation are yet to be solved. Methods: We have developed a method using two small molecules and no ectopic gene expression to develop patient-specific hepatic stem cells (hCdH) from primary hepatocytes. Of interest, from 52 patients healthy and diseased patient livers, we were able to generate hCdH. Results: Among the 52 patients, significant differences in fibrosis were observed with 4.17% and 52% in the normal and diseased group respectively. The harvested hepatocytes ratio (number of cells/the liver weight) also differed with a mean of 5.9x10⁶ in normal livers and 1.1x10⁶ in diseased livers. In normal livers ALP ranged from 66 to 95 U/L whilst there was a significant increase in the ALP level of diseased livers ranging from 80 to 174 U/L. Through the evaluation of different platforms such as generation and differentiation ratio, hepatic marker expression and functionality of these hCdHs, we were able to find the most significant difference in the statistical analysis between the generation of CdH in healthy and diseased livers and the level of ALP(alkaline phosphatase). Through FACS analysis we were also able to find the stemness marker NPB24-B3 obtained from embryonic stem cells as a characteristic of CdH and sorted 36.2% of the cells accordingly. Conclusions: Although the CdH characteristics vary, it was possible to generate CdH from all patients. We therefore suggest that the functional state and generation efficiency of CdH are related to patient liver condition. In further studies, we plan to use the CdH marker to sort the cells obtained from the patients to allow for homogeneous population.