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Article ; Online: Optimized conditions for gene transduction into primary immune cells using viral vectors

Yeongrin Kim / Da Yeon Lee / Ji U Choi / Jin Song Park / So Myoung Lee / Chung Hyo Kang / Chi Hoon Park

Scientific Reports, Vol 13, Iss 1, Pp 1-

2023 Volume 12

Abstract: Abstract Chimeric antigen receptor (CAR) T cell therapy has emerged as a promising modality for anti-cancer treatment. Its efficacy is quite remarkable in hematological tumors. Owing to their excellent clinical results, gene- modified cell therapies, ... ...

Abstract	Abstract Chimeric antigen receptor (CAR) T cell therapy has emerged as a promising modality for anti-cancer treatment. Its efficacy is quite remarkable in hematological tumors. Owing to their excellent clinical results, gene- modified cell therapies, including T cells, natural killer (NK) cells, and macrophages, are being actively studied in both academia and industry. However, the protocol to make CAR immune cells is too complicated, so it is still unclear how to efficiently produce the potent CAR immune cells. To manufacture effective CAR immune cells, we need to be aware of not only how to obtain highly infective viral particles, but also how to transduce CAR genes into immune cells. In this paper, we provide detailed information on spinoculation, which is one of the best known protocols to transduce genes into immune cells, in a methodological view. Our data indicate that gene transduction is significantly dependent on speed and duration of centrifugation, concentration and number of viral particles, the concentration of polybrene, and number of infected immune cells. In addition, we investigated on the optimal polyethylene glycol (PEG) solution to concentrate the viral supernatant and the optimized DNA ratios transfected into 293T cells to produce high titer of viral particles. This study provides useful information for practical production of the gene-modified immune cells using viral vectors.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2023-07-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Crystal structure of [1,2,4] triazolo[4,3- b ]pyridazine derivatives as BRD4 bromodomain inhibitors and structure–activity relationship study

Jung-Hoon Kim / Navin Pandit / Miyoun Yoo / Tae Hyun Park / Ji U Choi / Chi Hoon Park / Kwan-Young Jung / Byung Il Lee

Scientific Reports, Vol 13, Iss 1, Pp 1-

2023 Volume 12

Abstract: Abstract BRD4 contains two tandem bromodomains (BD1 and BD2) that recognize acetylated lysine for epigenetic reading, and these bromodomains are promising therapeutic targets for treating various diseases, including cancers. BRD4 is a well-studied target, ...

Abstract	Abstract BRD4 contains two tandem bromodomains (BD1 and BD2) that recognize acetylated lysine for epigenetic reading, and these bromodomains are promising therapeutic targets for treating various diseases, including cancers. BRD4 is a well-studied target, and many chemical scaffolds for inhibitors have been developed. Research on the development of BRD4 inhibitors against various diseases is actively being conducted. Herein, we propose a series of [1,2,4]triazolo[4,3-b]pyridazine derivatives as bromodomain inhibitors with micromolar IC50 values. We characterized the binding modes by determining the crystal structures of BD1 in complex with four selected inhibitors. Compounds containing [1,2,4] triazolo[4,3-b]pyridazine derivatives offer promising starting molecules for designing potent BRD4 BD inhibitors.
Keywords	Medicine ; R ; Science ; Q
Language	English
Publishing date	2023-07-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Identification of Potent CD19 scFv for CAR T Cells through scFv Screening with NK/T-Cell Line

Chung Hyo Kang / Yeongrin Kim / Heung Kyoung Lee / So Myoung Lee / Hye Gwang Jeong / Sang Un Choi / Chi Hoon Park

International Journal of Molecular Sciences, Vol 21, Iss 9163, p

2020 Volume 9163

Abstract: CD19 is the most promising target for developing chimeric-antigen receptor (CAR) T cells against B-cell leukemic cancer. Currently, two CAR-T-cell products, Kymriah and Yescarta, are approved for leukemia patients, and various anti-CD19 CAR T cells are ... ...

Abstract	CD19 is the most promising target for developing chimeric-antigen receptor (CAR) T cells against B-cell leukemic cancer. Currently, two CAR-T-cell products, Kymriah and Yescarta, are approved for leukemia patients, and various anti-CD19 CAR T cells are undergoing clinical trial. Most of these anti-CD19 CAR T cells use FMC63 single-chain variable fragments (scFvs) for binding CD19 expressed on the cancer cell surface. In this study, we screened several known CD19 scFvs for developing anti-CD19 CAR T cells. We used the KHYG-1 NK/T-cell line for screening of CD19 scFvs because it has advantages in terms of cell culture and gene transduction compared to primary T cells. Using our CAR construct backbone, we made anti-CD19 CAR constructs which each had CD19 scFvs including FMC63, B43, 25C1, BLY3, 4G7, HD37, HB12a, and HB12b, then made each anti-CD19 CAR KHYG-1 cells. Interestingly, only FMC63 CAR KHYG-1 and 4G7 CAR KHYG-1 efficiently lysed CD19-positive cell lines. In addition, in Jurkat cell line, only these two CAR Jurkat cell lines secreted IL-2 when co-cultured with CD19-positive cell line, NALM-6. Based on these results, we made FMC63 CAR T cells and 4G7 CAR T cells from PBMC. In in vitro lysis assay, 4G7 CAR T cells lysed CD19-positive cell line as well as FMC63 CAR T cells. In in vivo assay with NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice, 4G7 CAR T cells eradicated NALM-6 as potently as FMC63 CAR T cells. Therefore, we anticipate that 4G7 CAR T cells will show as good a result as FMC63 CAR T cells for B-cell leukemia patients.
Keywords	CD19 ; chimeric-antigen receptor ; scFv ; FMC63 ; 4G7 ; leukemia ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
Language	English
Publishing date	2020-12-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Supporting data for impact of filler composition on mechanical and dynamic response of 3-D printed silicone-based nanocomposite elastomers

Samantha J. Talley / Brittany Branch / Cynthia F. Welch / Chi Hoon Park / Dana M. Dattelbaum / Kwan-Soo Lee

Data in Brief, Vol 32, Iss , Pp 106240- (2020)

2020

Abstract: This research reports on the physical and mechanical effects of various filler materials used in direct ink write (DIW) 3-D printing resins. The data reported herein supports interpretation and discussion provided in the research article “Impact of ... ...

Abstract	This research reports on the physical and mechanical effects of various filler materials used in direct ink write (DIW) 3-D printing resins. The data reported herein supports interpretation and discussion provided in the research article “Impact of Filler Composition on Mechanical and Dynamic Response of 3-D Printed Silicone-based Nanocomposite Elastomers” [1]. The datasheet describes the model structures and the interaction energies between the fillers and the other components by using Molecular Dynamics (MD) simulations. This report includes mechanical responses of single-cubic (SC) and face-centered tetragonal (FCT) structures printed using new DIW resin formulations (polydimethylsiloxane-based silicones filled with aluminum oxide, graphite, or titanium dioxide). Using MD simulations and mechanical data, the overall flexibility and interactions between resin components are fully characterized.
Keywords	3-D printing ; Nanocomposite elastomer ; Molecular dynamics simulation ; Dynamic response ; Silicone ; Computer applications to medicine. Medical informatics ; R858-859.7 ; Science (General) ; Q1-390
Language	English
Publishing date	2020-10-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Synthesis and Structure–Activity Relationships of Aristoyagonine Derivatives as Brd4 Bromodomain Inhibitors with X-ray Co-Crystal Research

Minjin Yoo / Tae Hyun Park / Miyoun Yoo / Yeongrin Kim / Joo-Youn Lee / Kyu Myung Lee / Seong Eon Ryu / Byung Il Lee / Kwan-Young Jung / Chi Hoon Park

Molecules, Vol 26, Iss 1686, p

2021 Volume 1686

Abstract: Epigenetic regulation is known to play a key role in progression of anti-cancer therapeutics. Lysine acetylation is an important mechanism in controlling gene expression. There has been increasing interest in bromodomain owing to its ability to modulate ... ...

Abstract	Epigenetic regulation is known to play a key role in progression of anti-cancer therapeutics. Lysine acetylation is an important mechanism in controlling gene expression. There has been increasing interest in bromodomain owing to its ability to modulate transcription of various genes as an epigenetic ‘reader.’ Herein, we report the design, synthesis, and X-ray studies of novel aristoyagonine (benzo[6,7]oxepino[4,3,2- cd ]isoindol-2(1 H )-one) derivatives and investigate their inhibitory effect against Brd4 bromodomain. Five compounds 8ab , 8bc , 8bd , 8be , and 8bf have been discovered with high binding affinity over the Brd4 protein. Co-crystal structures of these five inhibitors with human Brd4 bromodomain demonstrated that it has a key binding mode occupying the hydrophobic pocket, which is known to be the acetylated lysine binding site. These novel Brd4 bromodomain inhibitors demonstrated impressive inhibitory activity and mode of action for the treatment of cancer diseases.
Keywords	Brd4 ; aristoyagonine ; epigenetic ; crystal structure ; SAR ; Organic chemistry ; QD241-441
Subject code	540
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Identification of novel ALK2 inhibitors and their effect on cancer cells

Kim, Minsung / Chi Hoon Park / Okran Choi / Sang Un Choi / Suhkneung Pyo

Biochemical and biophysical research communications. 2017 Oct. 07, v. 492, no. 1

2017

Abstract: Bone morphogenetic protein 9 (BMP9), a member of the TGF-β superfamily, is considered a regulator of glucose homeostasis as well as a neuronal differentiation factor. BMP9 induces phosphorylation of Smad1/5 through activin receptor-like kinase 1 and 2 ( ... ...

Abstract	Bone morphogenetic protein 9 (BMP9), a member of the TGF-β superfamily, is considered a regulator of glucose homeostasis as well as a neuronal differentiation factor. BMP9 induces phosphorylation of Smad1/5 through activin receptor-like kinase 1 and 2 (ALK1 and ALK2). Recently, many studies have shown that BMP9 contributes to tumorigenesis, and aberrant ALK2 expression is involved in many diseases. To investigate the role of BMP9-ALK2 signaling in cancer cells, we used TF-1 cells that require granulocyte-macrophage colony-stimulating factor (GM-CSF) for cell proliferation. BMP9 promoted the proliferation of TF-1 cells in media lacking GM-CSF. TF-1 cells overexpressing ALK2 resulted in the autophosphorylation of Smad1/5, leading to consequent increase in cell growth. Through high-throughput screening (HTS), we found two ALK2-specific inhibitors, KRC203 and KRC360, with IC50 values of 0.9 nM and 0.3 nM. These compounds were more potent and specific for the inhibition of ALK2 when compared to LDN193189. In cell-based assays, these compounds effectively inhibited the proliferation and migration of cancer cells induced by ALK2 and BMP9. Therefore, we propose that our compounds are promising candidates for the treatment of cancer or diseases with abnormal ALK2 or BMP9 signaling.
Keywords	bone morphogenetic proteins ; carcinogenesis ; cell growth ; cell proliferation ; glucose ; granulocyte-macrophage colony-stimulating factor ; homeostasis ; inhibitory concentration 50 ; neoplasm cells ; neoplasms ; neurons ; protein phosphorylation ; screening ; transforming growth factor beta
Language	English
Dates of publication	2017-1007
Size	p. 121-127.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2017.08.016
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Folate receptor 1 (FOLR1) targeted chimeric antigen receptor (CAR) T cells for the treatment of gastric cancer.

Minsung Kim / Suhkneung Pyo / Chung Hyo Kang / Chong Ock Lee / Heung Kyoung Lee / Sang Un Choi / Chi Hoon Park

PLoS ONE, Vol 13, Iss 6, p e

2018 Volume 0198347

Abstract: Gastric cancer is a malignancy that has a high mortality rate. Although progress has been made in the treatment of gastric cancer, many patients experience cancer recurrence and metastasis. Folate receptor 1 (FOLR1) is overexpressed on the cell surface ... ...

Abstract	Gastric cancer is a malignancy that has a high mortality rate. Although progress has been made in the treatment of gastric cancer, many patients experience cancer recurrence and metastasis. Folate receptor 1 (FOLR1) is overexpressed on the cell surface in over one-third of gastric cancer patients, but rarely is expressed in normal tissue. This makes FOLR1 a potential target for chimeric antigen receptor (CAR) T cell immunotherapy, although the function of FOLR1 has not been elucidated. CAR are engineered fusion receptor composed of an antigen recognition region and signaling domains. T cells expressing CAR have specific activation and cytotoxic effects against cancer cells containing the target antigen. In this study, we generated a CAR that targets FOLR1 composed of a single-chain variable fragment (scFv) of FOLR1 antibody and signaling domains consisting of CD28 and CD3ζ. Both FOLR1-CAR KHYG-1, a natural killer cell line, and FOLR1-CAR T cells recognized FOLR1-positive gastric cancer cells in a MHC-independent manner and induced secretion of various cytokines and caused cell death. Conclusively, this is the first study to demonstrate that CAR KHYG-1/T cells targeting FOLR1 are effective against FOLR1-positive gastric cancer cells.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2018-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Rapid custom prototyping of soft poroelastic biosensor for simultaneous epicardial recording and imaging

Bongjoong Kim / Arvin H. Soepriatna / Woohyun Park / Haesoo Moon / Abigail Cox / Jianchao Zhao / Nevin S. Gupta / Chi Hoon Park / Kyunghun Kim / Yale Jeon / Hanmin Jang / Dong Rip Kim / Hyowon Lee / Kwan-Soo Lee / Craig J. Goergen / Chi Hwan Lee

Nature Communications, Vol 12, Iss 1, Pp 1-

2021 Volume 14

Abstract: Printed biosensors are important for health monitoring and research purposes. Here, the authors report on the development of a soft poroelastic silicone based sensor which can be easily printed and is resistant to mechanical strain hysteresis, allowing ... ...

Abstract	Printed biosensors are important for health monitoring and research purposes. Here, the authors report on the development of a soft poroelastic silicone based sensor which can be easily printed and is resistant to mechanical strain hysteresis, allowing for more accurate electrophysiology readings and imaging.
Keywords	Science ; Q
Language	English
Publishing date	2021-06-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Sorption and Diffusion of CO2/N2 in gas mixture in thermally-rearranged polymeric membranes: A molecular investigation

Rizzuto, Carmen / Adele Brunetti / Alessio Caravella / Chi Hoon Park / Elena Tocci / Enrico Drioli / Giuseppe Barbieri / Young Moo Lee

Journal of membrane science. 2017 Apr. 15, v. 528

2017

Abstract: In this work, we study the adsorption and diffusion of nitrogen and carbon dioxide through an atomistically detailed model of a thermally rearranged polybenzoxazole (TR-PBO) polymer membranes, via equilibrium molecular dynamics (MD) simulations. This ... ...

Abstract	In this work, we study the adsorption and diffusion of nitrogen and carbon dioxide through an atomistically detailed model of a thermally rearranged polybenzoxazole (TR-PBO) polymer membranes, via equilibrium molecular dynamics (MD) simulations. This work represents a first explicit molecular modelling of the behavior of CO2/N2 binary mixture in TR-PBO and demonstrates how diffusivity and solubility in mixtures can be coherently obtained. In particular, the number of molecules present in the polymer matrix is estimated using the Gran Canonical Monte Carlo approach. As for the sorption in mixture conditions, MD simulations are used in a synergistic pairing with GCMC and Ideal Adsorption Solution Theory (IAST). For this purpose, the single-gas isotherms calculated from GCMC simulations are fitted with Langmuir and Dual-Langmuir adsorption models to obtain the parameters needed for the IAST simulations.As for diffusion, single-gas and mixture (Maxwell-Stefan) diffusion coefficients are performed by MD simulations. As main results, it is observed that the evaluated diffusion coefficients of CO2 and N2 are in a satisfactory agreement with the values estimated using the available experimental permeability data. More specifically, the CO2 diffusivity in mixture conditions is found to be the same as that in the single-gas one, whereas the N2 diffusivity is slightly higher. These differences are explained in terms of the effect of both the mutual gas diffusion and the competing occupancy of the available free space preferentially occupied by the CO2 molecules in mixture.
Keywords	adsorption ; artificial membranes ; carbon dioxide ; diffusivity ; molecular dynamics ; molecular models ; nitrogen ; permeability ; polymers ; solubility
Language	English
Dates of publication	2017-0415
Size	p. 135-146.
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	194516-6
ISSN	0376-7388
ISSN	0376-7388
DOI	10.1016/j.memsci.2017.01.025
Database	NAL-Catalogue (AGRICOLA)

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Article: Synthesis and biological evaluation of indazole-4,7-dione derivatives as novel BRD4 inhibitors

Yoo, Minjin / Miyoun Yoo / Ji Eun Kim / Heung Kyoung Lee / Chong Ock Lee / Chi Hoon Park / Kwan-Young Jung

Archives of pharmacal research. 2018 Jan., v. 41, no. 1

2018

Abstract: Bromodomain-containing protein 4 (BRD4) is known to regulate the expression of c-Myc to control the proliferation of cancer cells. Therefore, development of small-molecule inhibitors targeting the bromodomain has been widely studied. However, some ... ...

Abstract	Bromodomain-containing protein 4 (BRD4) is known to regulate the expression of c-Myc to control the proliferation of cancer cells. Therefore, development of small-molecule inhibitors targeting the bromodomain has been widely studied. However, some clinical trials on BRD4 inhibitors have shown its drawbacks such as toxicity including the loss of organ weight. Here, we report the development of the novel and promising scaffold, 1H-indazol-4,7-dione, as a bromodomain inhibitor and synthesized derivatives for the inhibition of binding of bromodomain to acetylated histone peptide. Through this effort, we obtained 6-chloro-5-((2,6-difluorophenyl)amino)-1H-indazole-4,7-dione (5i), which showed a highly potent activity with a half-maximal inhibitory concentration (IC₅₀) of 60 nM. The in vivo xenograft assay confirmed that the 1H-indazol-4,7-dione compound reduced the tumor size significantly. These results show that the 1H-indazol-4,7-dione scaffold is highly potent against bromodomain.
Keywords	clinical trials ; histones ; inhibitory concentration 50 ; neoplasm cells ; neoplasms ; tissue weight ; toxicity
Language	English
Dates of publication	2018-01
Size	p. 46-56.
Publishing place	Pharmaceutical Society of Korea
Document type	Article
ZDB-ID	447623-2
ISSN	1976-3786 ; 0253-6269
ISSN (online)	1976-3786
ISSN	0253-6269
DOI	10.1007/s12272-017-0978-y
Database	NAL-Catalogue (AGRICOLA)

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