Overexpression of histone methyltransferase NSD in <i>Drosophila</i> induces apoptotic cell death via the Jun-N-terminal kinase pathway

Jeong, Yuji,Kim, Taejoon,Kim, Suyeun,Hong, Yoon-Ki,Cho, Kyoung Sang,Lee, Im-Soon Elsevier 2018 Biochemical and biophysical research communication Vol.496 No.4

<P><B>Abstract</B></P> <P>The nuclear receptor-binding SET domain protein gene (<I>NSD</I>) family encodes a group of highly conserved SET domain-containing histone lysine methyltransferases that are important in multiple aspects of development in various organisms. The association of <I>NSD1</I> duplications has been reported with growth retardation diseases in humans. In this study, to gain insight into the molecular mechanisms by which the overexpression of <I>NSD1</I> influences the disease progression, we analyzed the gain-of-function mutant phenotypes of the <I>Drosophila NSD</I> using the <I>GAL4/UAS</I> system. Ubiquitous overexpression of <I>NSD</I> in the fly caused developmental delay and reduced body size at the larval stage, resulting in pupal lethality. Moreover, targeted overexpression in various developing tissues led to significant phenotype alterations, and the gain-of-function phenotypes were rescued by <I>NSD</I> RNAi knockdown. We also demonstrated that <I>NSD</I> overexpression not only enhanced the transcription of pro-apoptotic genes but also activated caspase. The atrophied phenotype of <I>NSD</I>-overexpressing wing was strongly suppressed by a loss-of-function mutation in <I>hemipterous</I>, which encodes a <I>Drosophila</I> Jun N-terminal kinase. Taken together, our findings suggest that NSD induces apoptosis via the activation of JNK, and thus contributes to the understanding of the molecular mechanisms involved in <I>NSD1</I>-related diseases in humans.</P> <P><B>Highlights</B></P> <P> <UL> <LI> NSD overexpression in <I>Drosophila</I> causes developmental delay and reduced body size. </LI> <LI> It resembles phenotypes of patients with <I>NSD1</I> duplication. </LI> <LI> NSD overexpression induces apoptosis in various imaginal discs of <I>Drosophila.</I> </LI> <LI> Apoptosis induced by NSD overexpression is suppressed by inhibiting the JNK pathway. </LI> </UL> </P>

Drosophila NSD deletion induces developmental anomalies similar to those seen in Sotos syndrome 1 patients

Saeyan Choi,Bokyeong Song,Hyewon Shin,Chihyun Won,Taejoon Kim,Hideki Yoshida,Daewon Lee,Jongkyeong Chung,Kyoung Sang Cho,이임순 한국유전학회 2021 Genes & Genomics Vol.43 No.7

Background Haploinsufciency of the human nuclear receptor binding suppressor of variegation 3–9, enhancer of zeste, and trithorax (SET) domain 1 (NSD1) gene causes a developmental disorder called Sotos syndrome 1 (SOTOS1), which is associated with overgrowth and macrocephaly. NSD family proteins encoding histone H3 lysine 36 (H3K36) methyltransferases are conserved in many species, and Drosophila has a single NSD homolog gene, NSD. Objective To gain insight into the biological functions of NSD1 defciency in the developmental anomalies seen in SOTOS1 patients using an NSD-deleted Drosophila mutant. Methods We deleted Drosophila NSD using CRISPR/Cas9-mediated targeted gene knock-out, and analyzed pleiotropic phenotypes of the homozygous mutant of NSD (NSD−/−) at various developmental stages to understand the roles of NSD in Drosophila. Results The site-specifc NSD deletion was confrmed in the mutant. The H3K36 di-methylation levels were dramatically decreased in the NSD−/− fy. Compared with the control, the NSD−/− fy displayed an increase in the body size of larvae, similar to the childhood overgrowth phenotype of SOTOS1 patients. Although the NSD mutant fies survived to adulthood, their fecundity was dramatically decreased. Furthermore, the NSD−/− fy showed neurological dysfunctions, such as lower memory performance and motor defects, and a diminished extracellular signal-regulated kinase (ERK) activity. Conclusions The NSD-deleted Drosophila phenotype resembles many of the phenotypes of SOTOS1 patients, such as learning disability, deregulated ERK signaling, and overgrowth; thus, this mutant fy is a relevant model organism to study various SOTOS1 phenotypes.

Identification of LEM-14 inhibitor of the oncoprotein NSD2

Shen, Yunpeng,Morishita, Masayo,Lee, Doohyun,Kim, Shinae,Lee, Taeho,Mevius, Damiaan E.H.F.,Roh, Yeonjeong,di Luccio, Eric Elsevier 2019 Biochemical and biophysical research communication Vol.508 No.1

<P><B>Abstract</B></P> <P>The NSD family (NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L1) are histone lysine methyltransferases (HMTases) essential for chromatin regulation. The NSDs are oncoproteins, drivers of a number of tumors and are considered important drug-targets but the lack of potent and selective inhibitors hampers further therapeutic development and limits exploration of their biology. In particular, MMSET/NSD2 selective inhibition is being pursued for therapeutic interventions against multiple myeloma (MM) cases, especially in multiple myeloma t(4;14)(p16.3;q32) translocation that is associated with a significantly worse prognosis than other MM subgroups. Multiple myeloma is the second most common hematological malignancy, after non-Hodgkin lymphoma and remains an incurable malignancy.</P> <P>Here we report the discovery of LEM-14, an NSD2 specific inhibitor with an <I>in vitro</I> IC<SUB>50</SUB> of 132 μM and that is inactive against the closely related NSD1 and NSD3. LEM-14-1189, a LEM-14 derivative, differentially inhibits the NSDs with <I>in vitro</I> IC<SUB>50</SUB> of 418 μM (NSD1), IC<SUB>50</SUB> of 111 μM (NSD2) and IC<SUB>50</SUB> of 60 μM (NSD3). We propose LEM-14 and derivative LEM-14-1189 as tools for studying the biology of the NSDs and constitute meaningful steps toward potent NSDs therapeutic inhibitors.</P> <P><B>Highlights</B></P> <P> <UL> <LI> NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L1 are histone methyltransferases and oncoproteins. </LI> <LI> Inhibition of NSD2 in multiple myeloma t(4;14)(p16.3;q32) is urgently needed. </LI> <LI> We report the discovery of a NSD2 specific inhibitor and derivatives that differentially inhibits the NSDs. </LI> <LI> We investigate the molecular mechanism for NSD2 specific inhibition. </LI> <LI> We propose our inhibitors LEM-14 and LEM-14-1189 as tools for studying the biology of the NSDs and for drug-design. </LI> </UL> </P>

Structural Insight into the Post-SET Loop Conformation of NSD1 Implications for the Epigenetic Therapy of Cancers Targeting Histone Lysine Methyltransferases

디 루시오, 에릭 대한암예방학회 2011 Journal of cancer prevention Vol.16 No.2

Both genetic and epigenetic alterations are responsible for the stepwise initiation and progression of cancers. Only epigenetic aberrations can be reversible, allowing the malignant cell population to revert to a more benign phenotype. The epigenetic therapy of cancers is emerging as an effective and valuable approach to both chemotherapy and the chemoprevention of cancer. The utilization of epigenetic targets that include histone methyltransferase (HMTase), histone deacetylase, and DNA methyltransferase, are emerging as key therapeutic targets. The nuclear receptor binding SET domain (NSD) protein is a family of three HMTases, NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L1, and plays a critical part in chromatin integrity as evidenced by a growing number of conditions linked to the alterations or amplification of NSD1, NSD2, or NSD3. NSD1, NSD2 and NSD3 are associated with multiple cancers. The amplification of either NSD1 or NSD2 triggers the cellular transformation and thus is key in the early carcinogenesis events. In most cases, reducing the levels of NSDs would suppress cancer growth. The NSD pathways, however, are not well understood. Recently, a partial crystal structure of the ligandless SET domain of NSD1 has been published enabling further studies toward a structure-based drug design effort on the NSD family. Here, we use computational methods to model the binding of the natural ligand histone H4K20 on the SET domain of NSD1. Our findings unravel a critical insight into an autoregulatory mechanism driven by the flexibility of the post-SET loop region and we prospect its special value for developing novel anticancer drugs. (Cancer Prev Res 16, 93-101, 2011)

패스(Path)제어 방법을 통한 NSD 소음 저감 방안에 관한 연구

김선진(Seon-Jin Kim),김성곤(Sung-Gon Kim),강태우(Tae-Woo Kang),신철호(Cheol-Ho Shin) 한국산학기술학회 2018 한국산학기술학회논문지 Vol.19 No.10

본 연구는 패스(Path) 제어방법을 적용하여 소형전술차량에 적용된 NSD의 소음을 저감하기 위한 것이다. 소형전술차량은 고기동성 확보를 위해 후차축에 차동제한장치 중 하나인 NSD를 적용하였다. NSD는 높은 제한율을 가져 기동성을 향상 시켜주지만 기계적 구조에 의해 특정 조건에서 소음이 발생되는 단점이 있다. 이 소음은 NSD가 차동장치로써 원활한 역할수행을 위해 기어간 유격에 의해 치간 접촉에 따라 발생되는 소음이다. 이러한 소음이 지속적으로 사용자에게 전달됨에 따라 운용자가 지속적인 문제 제기 및 개선 요구를 하고 있다. 물론, NSD의 소음으로 인해 제품의 성능 또는 내구성에 미치는 영향은 없으며, 국방규격에서 규정하는 소음 조건 역시 만족한다. 하지만, 사용자의 지속적인 개선요구에 따라 소음저감을 위한 방안에 대해 연구를 수행하였다. 따라서, 본 연구에서는 소음원의 제거가 현실적으로 제한되는 전술차량의 NSD 소음을 감소하기 위해 NSD의 소음이 전달되는 경로를 제어하는 패스제어방법을 적용하였다. 이를 통해 구조 전달계를 개선•보완하고 이에 대한 개선효과를 검증하고자 한다. This paper presents means of reducing noise in NSD using path control methods for Light Tactical Vehicles (LTV). NSD is applied to the rear axle of LTVs for enhancing mobility. NSD can improve mobility of vehicles with a high locking ratio but causes noise under certain conditions due to its mechanical structure. This noise results from contact between gears due to the differential role of NSD. The noise affects users, so users have continually requested noise reductions. Though the noise doesn"t affect product performance and durability, and satisfies the National Defense"s noise condition standards, users request for noise reduction is the focus of this research. Eliminating the source of sound for LTVs is realistically limited, so this research applies a path control method to reduce noise by controlling the path which transmits the noise. This study improves the structural delivery system and examines methods of reducing noise in LTV systems.

고객지향적 서비스 상품 개발: 하나은행 사례

김병도 ( Byung Do Kim ),전종근 ( Jong Kun Jun ) 한국소비자학회 2011 소비자학연구 Vol.22 No.2

본 연구는 금융기관의 신서비스 개발 역량과 성과를 사례 연구를 통하여 분석하였다. 문헌연구를 통해 성공적인 신서비스 개발 프로세스의 요건을 파악하고 하나은행의 사례에서 이러한 요건들이 어떻게 나타나는지 비교 분석하였다. 하나은행은 공식적인 개발 프로세스의 보유, 테스트 마케팅 생략을 통한 출시시간 단축, 상업화 이후의 상품 리뷰를 통한 고객의 반응 추적 및 상품 보완 등에서 강점이 있었다. 반면, 고객을 신서비스 개발 프로세스에 적극적으로 참여시키는 부분, 위험을 감수하고 혁신을 장려하는 조직문화, 정보기술에 의존하는 신서비스 개발 프로세스 등에서는 개선이 필요한 것으로 파악되었다. 하나은행은 이러한 공식적인 신서비스 개발 프로세스와 함께 유연하고 자유로운 조직문화, 영업부서와 상품개발부서간의 긴밀한 협력을 결합하여 신서비스 개발 경쟁력을 강화하였다. 아울러 하나은행이 최근에 개발한 예금상품들의 경쟁력을 절충형 상품에 대한 소비자반응이라는 관점에서 분석한 결과, 첫째, 기능적 요소와 쾌락적 요소의 성공적 결합으로 인한 타협효과가 나타났을 가능성, 둘째, 금리변동기에 나타나는 상황적 방어초점이 절충형 상품에 대한 소비자의 긍정적 반응을 유발하였을 가능성 등을 파악하였으며 그에 따른 시사점을 제시하였다. This paper presents a case study on the new service development competence and performance of a bank. The authors compared the new service development(NSD) process of Hana Bank with the recommended NSD practices derived from literature review. Hana bank`s NSD process shares a few success factors with Alam and Perry(2001)`s customer-oriented new service development process such as focusing on idea generation and screening, following a sequential development process, and skipping test marketing to save time to market. Hana Bank showed its NSD competence by having a formal NSD process, omitting test marketing for a quick launch, and modifying its service followed by tracking customer response after launch. There were some practices that Hana Bank was not up to the mark including participating customers into the NSD process, cultivating an innovative and risk taking organizational culture, and utilizing IT throughout the NSD process. Hana Bank accomplished its NSD competence by combining its formal NSD process with a flexible organizational culture and close cooperation between functional departments. The Hana 369 Fixed Deposit is an innovative new savings product, which allows its customers to withdraw their money at every three months without penalty. It appealed to the consumers who wanted to have a little flexible deposit plan because they are uncertain about when they needed the money back. The new savings product has been very successful since it was launched in July, 2009. Another new saving product is S-Line Installment Savings, which offers additional interests to the customers who lost their weights successfully on its due day. These products have been successful because they both reflect the social and economic trends or situations. Another possible explanation for the success of the products is the compromise effect. Compromise alternative is preferred when people seek loss aversion, because its gains and losses are both smaller than those of other alternatives (Simonson and Tversky, 1992). The Hana 369 Fixed Deposit is a compromise alternative compared with one year fixed deposit plans and MMDAs. When the interest rates are rising, most people would not put their money in the fixed deposit plans for fear of opportunity loss. Especially, those people with prevention focus will likely to choose the Hana 369 Fixed Deposit over other alternatives since they want to minimize their losses. The principle of hedonic dominance is also useful to explain the consumer preference for the compromise alternative with hedonic elements such as the S-Line Installment Savings. Consumers prefer products with hedonic elements when both the hedonic and functional elements are above cutoff levels (Chitturi, Raghunathan, and Mahajan, 2007). Applying the theory to the S-Line Installment Savings, the hedonic element of going on a diet when you open an account will be attractive only when the interest rate is above cutoff level. The company will need to figure out what is the cutoff level of interest rate when people are planning to open a savings account. Additionally, trendy products are subject to the changes of the trends. The task of new service development department should be inclusive of continuous modification of products based on the monitoring of the trend changes. Based on the case study of Hana Bank the authors discussed the directions for the future NSD.

Recombinant Protein Expression and Purification of the Human HMTase MMSET/NSD2

Morishita, Masayo,Mevius, Damiaan,Shen, Yunpeng,Di Luccio, Eric Institute of Agricultural Science and Technology 2013 慶北大農學誌 Vol.31 No.3

Chromatin remodelers that include histone methyl transferases (HMTases) are becoming a focal point in cancer drug development. The NSD family of three HMTases, NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L are bona fide oncogenes found aberrantly expressed in several cancers, suggesting their potential role for novel therapeutic strategies. Several histone modifiers including HMTase have clear roles in human carcinogenesis but the extent of their functions and regulations are not well understood, especially in pathological conditions. The extents of the NSDs biological roles in normal and pathological conditions remain unclear. In particular, the substrate specificity of the NSDs remains unsettled and discrepant data has been reported. NSD2/MMSET is a focal point for therapeutic interventions against multiple myeloma and especially for t(4;14) myeloma, which is associated with a significantly worse prognosis than other biological subgroups. Multiple myeloma is the second most common hematological malignancy in the United States, after non-Hodgkin lymphoma. Herein, as a first step before entering a pipeline for protein x-ray crystallography, we cloned, recombinantly expressed and purified the catalytic SET domain of NSD2. Next, we demonstrated the catalytic activities, in vitro, of the recombinantly expressed NSD2-SET on H3K36 and H4K20, its biological targets at the chromatin.

Recombinant Protein Expression and Purification of the Human HMTase MMSET/NSD2

Masayo Morishita,Damiaan Mevius,Yunpeng Shen,Eric di Luccio 경북대학교 농업생명과학대학 2013 Current Research on Agriculture and Life Sciences Vol.31 No.3

Chromatin remodelers that include histone methyl transferases (HMTases) are becoming a focal point in cancer drug development. The NSD family of three HMTases, NSD1, NSD2/MMSET/WHSC1, and NSD3/WHSC1L are bona fide oncogenes found aberrantly expressed in several cancers, suggesting their potential role for novel therapeutic strategies. Several histone modifiers including HMTase have clear roles in human carcinogenesis but the extent of their functions and regulations are not well understood, especially in pathological conditions. The extents of the NSDs biological roles in normal and pathological conditions remain unclear. In particular, the substrate specificity of the NSDs remains unsettled and discrepant data has been reported. NSD2/MMSET is a focal point for therapeutic interventions against multiple myeloma and especially for t(4;14) myeloma, which is associated with a significantly worse prognosis than other biological subgroups. Multiple myeloma is the second most common hematological malignancy in the United States, after non-Hodgkin lymphoma. Herein, as a first step before entering a pipeline for protein x-ray crystallography, we cloned, recombinantly expressed and purified the catalytic SET domain of NSD2. Next, we demonstrated the catalytic activities, in vitro, of the recombinantly expressed NSD2-SET on H3K36 and H4K20, its biological targets at the chromatin.

NSD와 TDF에 관한 물리적 고찰

김성규,신세원,김명세 한국의학물리학회 1990 의학물리 Vol.1 No.1

On the basis of the review of radiobiological date, a formalism is developed for the analysis and prediction of iso-effect relations for tissue tolerance, which can be used as an alternative to the norminal standard dose(NSD) formaula of Ellis and its derived equations. An important feature of the described formalism is that directly based on radiobiological insights and it provides a more logical concept to account for the diversity of tissue responses. The NSD concept has subsequently been extended to the formalisms of timedose-fractionation(TDF) value. The authors deriveded TDF equation on the basis NSD of Ellis. TDF=0.07(NSD)-26.

Inhibition of Nuclear Receptor Binding SET Domain 2/ Multiple Myeloma SET Domain by LEM-06 Implication for Epigenetic Cancer Therapies

디 루시오, 에릭 대한암예방학회 2015 Journal of cancer prevention Vol.20 No.2

Background:Multiple myeloma SET domain (MMSET)/nuclear receptor binding SET domain 2 (NSD2) is a lysine histone methyltransferase (HMTase) and bona fideoncoprotein found aberrantly expressed in several cancers, suggesting potential role for novel therapeutic strategies. In particular, MMSET/NSD2 is emerging as a target for therapeutic interventions against multiple myeloma, especially t(4;14) myeloma that is associated with a significantly worse prognosis than other biological subgroups. Multiple myeloma is the secondmost common hematological malignancy in the United States, after non-Hodgkin lymphoma and remains an incurable malignancy. Thus, effective therapeutic strategies are greatly needed. HMTases inhibitors are scarce and no NSDs inhibitors have been isolated. Methods:We used homology modeling, molecular modeling simulations, virtual ligand screening, computational chemistry software for structure-activity relationship and performed in vitro H3K36 histone lysine methylation inhibitory assay using recombinant human NSD2-SET and human H3.1 histone. Results:Here, we report the discovery of LEM-06, a hit small molecule inhibitor of NSD2, with an IC50of 0.8 mM against H3K36 methylation in vitro. Conclusions:We propose LEM-06 as a hit inhibitor that is useful to further optimize for exploring the biology of NSD2. LEM-06 derivatives may pave the way to specific NSD2 inhibitors suitable for therapeutic efforts against malignancies.