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Article ; Online: A Comprehensive Review on the Role of ZSCAN4 in Embryonic Development, Stem Cells, and Cancer.

Thool, Madhuri / Sundaravadivelu, Pradeep Kumar / Sudhagar, S / Thummer, Rajkumar P

2022 Volume 18, Issue 8, Page(s) 2740–2756

Abstract: ZSCAN4 is a transcription factor that plays a pivotal role during early embryonic development. It is a unique gene expressed specifically during the first tide of de novo transcription during the zygotic genome activation. Moreover, it is reported to ... ...

Abstract	ZSCAN4 is a transcription factor that plays a pivotal role during early embryonic development. It is a unique gene expressed specifically during the first tide of de novo transcription during the zygotic genome activation. Moreover, it is reported to regulate telomere length in embryonic stem cells and induced pluripotent stem cells. Interestingly, ZSCAN4 is expressed in approximately 5% of the embryonic stem cells in culture at any given time, which points to the fact that it has a tight regulatory system. Furthermore, ZSCAN4, if included in the reprogramming cocktail along with core reprogramming factors, increases the reprogramming efficiency and results in better quality, genetically stable induced pluripotent stem cells. Also, it is reported to have a role in promoting cancer stem cell phenotype and can prospectively be used as a marker for the same. In this review, the multifaceted role of ZSCAN4 in embryonic development, embryonic stem cells, induced pluripotent stem cells, cancer, and germ cells are discussed comprehensively.
MeSH term(s)	Embryonic Development/genetics ; Embryonic Stem Cells/metabolism ; Gene Expression Regulation ; Induced Pluripotent Stem Cells ; Neoplasms/genetics ; Neoplasms/therapy ; Transcription Factors/genetics ; Transcription Factors/metabolism
Chemical Substances	Transcription Factors ; ZSCAN4 protein, human
Language	English
Publishing date	2022-06-23
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2495577-2
ISSN	2629-3277 ; 1558-6804 ; 1550-8943
ISSN (online)	2629-3277 ; 1558-6804
ISSN	1550-8943
DOI	10.1007/s12015-022-10412-1
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Zs.A 6198: Show issues

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Article ; Online: Tamoxifen modulates mitochondrial dynamics through AMPK and MAPK during nutrition deprivation.

Vijayakumar, Gangipangi / Swetha, Uppalapati S / Sudhagar, Selvaraju

Cell biology international

2022 Volume 46, Issue 10, Page(s) 1661–1671

Abstract: The interaction of cancer cells with their tumor microenvironment determines key events in the progression of the disease, therapeutic efficacy, and the development of drug resistance. Here, we presented evidence that tamoxifen support breast cancer ... ...

Abstract	The interaction of cancer cells with their tumor microenvironment determines key events in the progression of the disease, therapeutic efficacy, and the development of drug resistance. Here, we presented evidence that tamoxifen support breast cancer growth during nutrition deprivation by modulating mitochondrial dynamics through AMPK and MAPK signaling. Tamoxifen enhances mitochondrial fusion under nutrition-deprived conditions by suppressing Drp1 ser616 phosphorylation and upregulating Mfn1 levels. Tamoxifen-induced mitochondrial fusion is mediated by the activation of AMPK as evident by the pharmacological inhibition of AMPK reverse mitochondrial fusion. Interestingly, JNK activation by tamoxifen controls the mitochondrial fusion morphology by downregulating Mfn2. Collectively, tamoxifen support cell growth by enhancing mitochondrial fusion by regulating stress kinase signaling under nutrition deprivation condition.
MeSH term(s)	AMP-Activated Protein Kinases/metabolism ; Humans ; Mitochondrial Dynamics/physiology ; Phosphorylation ; Signal Transduction ; Tamoxifen/pharmacology
Chemical Substances	Tamoxifen (094ZI81Y45) ; AMP-Activated Protein Kinases (EC 2.7.11.31)
Language	English
Publishing date	2022-07-12
Publishing country	England
Document type	Journal Article
ZDB-ID	1143453-3
ISSN	1095-8355 ; 1065-6995
ISSN (online)	1095-8355
ISSN	1065-6995
DOI	10.1002/cbin.11853
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Zs.A 1451: Show issues

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Article: Generation of biologically active recombinant human OCT4 protein from

Dey, Chandrima / Thool, Madhuri / Bhattacharyya, Srirupa / Sudhagar, S / Thummer, Rajkumar P

3 Biotech

2021 Volume 11, Issue 5, Page(s) 207

Abstract: Octamer-binding transcription factor 4 (OCT4) is vital for early embryonic development and is a master regulator of pluripotency in embryonic stem cells. Notably, OCT4 is a key reprogramming factor to derive induced pluripotent stem cells, which have ... ...

Abstract	Octamer-binding transcription factor 4 (OCT4) is vital for early embryonic development and is a master regulator of pluripotency in embryonic stem cells. Notably, OCT4 is a key reprogramming factor to derive induced pluripotent stem cells, which have tremendous prospects in regenerative medicine. In the current study, we report heterologous expression and purification of human OCT4 in Supplementary information: The online version contains supplementary material available at 10.1007/s13205-021-02758-z.
Language	English
Publishing date	2021-04-08
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2600522-0
ISSN	2190-5738 ; 2190-572X
ISSN (online)	2190-5738
ISSN	2190-572X
DOI	10.1007/s13205-021-02758-z
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Article ; Online: An Insight into the Role of UTF1 in Development, Stem Cells, and Cancer.

Raina, Khyati / Dey, Chandrima / Thool, Madhuri / Sudhagar, S / Thummer, Rajkumar P

Stem cell reviews and reports

2021 Volume 17, Issue 4, Page(s) 1280–1293

Abstract: The curiosity to understand the mechanisms regulating transcription in pluripotent cells resulted in identifying a unique transcription factor named Undifferentiated embryonic cell transcription factor 1 (UTF1). This proline-rich, nuclear protein is ... ...

Abstract	The curiosity to understand the mechanisms regulating transcription in pluripotent cells resulted in identifying a unique transcription factor named Undifferentiated embryonic cell transcription factor 1 (UTF1). This proline-rich, nuclear protein is highly conserved among placental mammals with prominent expression observed in pluripotent, germ, and cancer cells. In pluripotent and germ cells, its role has been implicated primarily in proper cell differentiation, whereas in cancer, it shows tissue-specific function, either as an oncogene or a tumor suppressor gene. Furthermore, UTF1 is crucial for germ cell development, spermatogenesis, and maintaining male fertility in mice. In addition, recent studies have demonstrated the importance of UTF1 in the generation of high quality induced Pluripotent Stem Cells (iPSCs) and as an excellent biomarker to identify bona fide iPSCs. Functionally, UTF1 aids in establishing a favorable chromatin state in embryonic stem cells, reducing "transcriptional noise" and possibly functions similarly in re-establishing this state in differentiated cells upon their reprogramming to generate mature iPSCs. This review highlights the multifaceted roles of UTF1 and its implication in development, spermatogenesis, stem, and cancer cells.
MeSH term(s)	Animals ; Chromosomal Proteins, Non-Histone/physiology ; Embryonic Stem Cells ; Female ; Male ; Mice ; Neoplasms/genetics ; Nuclear Proteins ; Placenta ; Pregnancy ; Trans-Activators/physiology ; Transcription Factors
Chemical Substances	Chromosomal Proteins, Non-Histone ; Nuclear Proteins ; Trans-Activators ; Transcription Factors ; Utf1 protein, mouse
Language	English
Publishing date	2021-01-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2495577-2
ISSN	2629-3277 ; 1558-6804 ; 1550-8943
ISSN (online)	2629-3277 ; 1558-6804
ISSN	1550-8943
DOI	10.1007/s12015-021-10127-9
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Zs.A 6198: Show issues

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Article ; Online: Generation of a Recombinant Stem Cell-Specific Human SOX2 Protein from Escherichia coli Under Native Conditions.

Thool, Madhuri / Dey, Chandrima / Bhattacharyya, Srirupa / Sudhagar, S / Thummer, Rajkumar P

Molecular biotechnology

2021 Volume 63, Issue 4, Page(s) 327–338

Abstract: The stem cell-specific SOX2 transcription factor is critical for early embryonic development and the maintenance of embryonic and neural stem cell identity. It is also crucial for the generation of induced pluripotent and neural stem cells, thus ... ...

Abstract	The stem cell-specific SOX2 transcription factor is critical for early embryonic development and the maintenance of embryonic and neural stem cell identity. It is also crucial for the generation of induced pluripotent and neural stem cells, thus providing immense prospect in patient-specific therapies. Here, we report soluble expression and purification of human SOX2 protein under native conditions from a bacterial system. To generate this macromolecule, we codon-optimized the protein-coding sequence and fused it to a nuclear localization signal, a protein transduction domain, and a His-tag. This was then cloned into a protein expression vector and was expressed in Escherichia coli. Subsequently, we have screened and identified the optimal expression conditions to obtain recombinant fusion protein in a soluble form and studied its expression concerning the position of fusion tags at either terminal. Furthermore, we purified two versions of recombinant SOX2 fusion proteins to homogeneity under native conditions and demonstrated that they maintained their secondary structure. This molecular tool can substitute genetic and viral forms of SOX2 to facilitate the derivation of integration-free induced pluripotent and neural stem cells. Furthermore, it can be used in elucidating its role in stem cells, various cellular processes and diseases, and for structural and biochemical studies.
MeSH term(s)	Chromatography, Affinity ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/growth & development ; Escherichia coli/metabolism ; Humans ; Models, Molecular ; Nuclear Localization Signals ; Protein Engineering ; Protein Structure, Secondary ; Recombinant Fusion Proteins/metabolism ; SOXB1 Transcription Factors/genetics ; SOXB1 Transcription Factors/metabolism ; Stem Cells/metabolism
Chemical Substances	Nuclear Localization Signals ; Recombinant Fusion Proteins ; SOX2 protein, human ; SOXB1 Transcription Factors
Language	English
Publishing date	2021-02-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	1193057-3
ISSN	1559-0305 ; 1073-6085
ISSN (online)	1559-0305
ISSN	1073-6085
DOI	10.1007/s12033-021-00305-y
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Zs.A 4031: Show issues

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Article: Tissue-Restricted Stem Cells as Starting Cell Source for Efficient Generation of Pluripotent Stem Cells: An Overview.

Sundaravadivelu, Pradeep Kumar / Raina, Khyati / Thool, Madhuri / Ray, Arnab / Joshi, Jahnavy Madhukar / Kaveeshwar, Vishwas / Sudhagar, S / Lenka, Nibedita / Thummer, Rajkumar P

Advances in experimental medicine and biology

2021 Volume 1376, Page(s) 151–180

Abstract: Induced pluripotent stem cells (iPSCs) have vast biomedical potential concerning disease modeling, drug screening and discovery, cell therapy, tissue engineering, and understanding organismal development. In the year 2006, a groundbreaking study reported ...

Abstract	Induced pluripotent stem cells (iPSCs) have vast biomedical potential concerning disease modeling, drug screening and discovery, cell therapy, tissue engineering, and understanding organismal development. In the year 2006, a groundbreaking study reported the generation of iPSCs from mouse embryonic fibroblasts by viral transduction of four transcription factors, namely, Oct4, Sox2, Klf4, and c-Myc. Subsequently, human iPSCs were generated by reprogramming fibroblasts as a starting cell source using two reprogramming factor cocktails [(i) OCT4, SOX2, KLF4, and c-MYC, and (ii) OCT4, SOX2, NANOG, and LIN28]. The wide range of applications of these human iPSCs in research, therapeutics, and personalized medicine has driven the scientific community to optimize and understand this reprogramming process to achieve quality iPSCs with higher efficiency and faster kinetics. One of the essential criteria to address this is by identifying an ideal cell source in which pluripotency can be induced efficiently to give rise to high-quality iPSCs. Therefore, various cell types have been studied for their ability to generate iPSCs efficiently. Cell sources that can be easily reverted to a pluripotent state are tissue-restricted stem cells present in the fetus and adult tissues. Tissue-restricted stem cells can be isolated from fetal, cord blood, bone marrow, and other adult tissues or can be obtained by differentiation of embryonic stem cells or trans-differentiation of other tissue-restricted stem cells. Since these cells are undifferentiated cells with self-renewal potential, they are much easier to reprogram due to the inherent characteristic of having an endogenous expression of few pluripotency-inducing factors. This review presents an overview of promising tissue-restricted stem cells that can be isolated from different sources, namely, neural stem cells, hematopoietic stem cells, mesenchymal stem cells, limbal epithelial stem cells, and spermatogonial stem cells, and their reprogramming efficacy. This insight will pave the way for developing safe and efficient reprogramming strategies and generating patient-specific iPSCs from tissue-restricted stem cells derived from various fetal and adult tissues.
MeSH term(s)	Animals ; Cell Differentiation ; Cells, Cultured ; Cellular Reprogramming ; Embryonic Stem Cells ; Fibroblasts/metabolism ; Humans ; Induced Pluripotent Stem Cells ; Kruppel-Like Factor 4 ; Mice ; Octamer Transcription Factor-3/genetics ; Octamer Transcription Factor-3/metabolism
Chemical Substances	Kruppel-Like Factor 4 ; Octamer Transcription Factor-3
Language	English
Publishing date	2021-10-06
Publishing country	United States
Document type	Journal Article ; Review
ISSN	2214-8019 ; 0065-2598
ISSN (online)	2214-8019
ISSN	0065-2598
DOI	10.1007/5584_2021_660
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Article: Inhibition of epidermal growth factor receptor by ferulic acid and 4-vinylguaiacol in human breast cancer cells

Sudhagar, S / S. Sathya / R. Anuradha / G. Gokulapriya / Y. Geetharani / B. S. Lakshmi

Biotechnology letters. 2018 Feb., v. 40, no. 2

2018

Abstract: OBJECTIVES: To examine the potential of ferulic acid and 4-vinylguaiacol for inhibiting epidermal growth factor receptor (EGFR) in human breast cancer cells in vitro. RESULTS: Ferulic acid and 4-vinylguaiacol limit the EGF (epidermal growth factor)- ... ...

Abstract	OBJECTIVES: To examine the potential of ferulic acid and 4-vinylguaiacol for inhibiting epidermal growth factor receptor (EGFR) in human breast cancer cells in vitro. RESULTS: Ferulic acid and 4-vinylguaiacol limit the EGF (epidermal growth factor)-induced breast cancer proliferation and new DNA synthesis. Western blot analysis revealed both ferulic acid and 4-vinylguaiacol exhibit sustained inhibition of EGFR activation through down-regulation of Tyr 1068 autophosphorylation. Molecular docking analysis shows ferulic acid forming hydrogen bond interaction with Lys 745 and Met 793 whereas, 4-vinylguaiacol forms two hydrogen bonds with Phe 856 and exhibits stronger hydrophobic interactions with multiple amino acid residues at the EGFR kinase domain. CONCLUSIONS: Ferulic acid and 4-vinylguaiacol could serve as a potential structure for the development of new small molecule therapeutics against EGFR.
Keywords	DNA replication ; Western blotting ; amino acids ; breast neoplasms ; epidermal growth factor ; epidermal growth factor receptors ; ferulic acid ; humans ; hydrogen bonding ; hydrophobic bonding ; molecular models ; neoplasm cells ; protein phosphorylation ; therapeutics
Language	English
Dates of publication	2018-02
Size	p. 257-262.
Publishing place	Springer Netherlands
Document type	Article
ZDB-ID	423853-9
ISSN	1573-6776 ; 0141-5492
ISSN (online)	1573-6776
ISSN	0141-5492
DOI	10.1007/s10529-017-2475-2
Database	NAL-Catalogue (AGRICOLA)

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Zs.A 4564: Show issues

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Article ; Online: Investigation of ROS generating capacity of curcumin-loaded liposomes and its in vitro cytotoxicity on MCF-7 cell lines using photodynamic therapy.

Prathyusha, Eluri / A, Prabakaran / Ahmed, Hafiz / Dethe, Mithun Rajendra / Agrawal, Mukta / Gangipangi, Vijayakumar / Sudhagar, S / Krishna, Kowthavarapu Venkata / Dubey, Sunil Kumar / Pemmaraju, Deepak B / Alexander, Amit

Photodiagnosis and photodynamic therapy

2022 Volume 40, Page(s) 103091

Abstract: Photodynamic therapy (PDT) is highly efficient in eradicating targetlesions by using photosensitizers (PS) triggered by external light energy. Nanotechnology may help increase the solubility and effective delivery of PS towards improving its efficacy. ... ...

Abstract	Photodynamic therapy (PDT) is highly efficient in eradicating targetlesions by using photosensitizers (PS) triggered by external light energy. Nanotechnology may help increase the solubility and effective delivery of PS towards improving its efficacy. Curcumin (Cur) was used as a natural PS for PDT in the present work. Briefly, curcumin was encapsulated in liposomes (LPs) using the thin film hydration method and optimized using the QbD approach through the Box-Behnken Design (BBD) to optimize the responses like entrapment efficiency and drug loading with a smaller vesicle size. The in vitro release studies performed using a dialysis bag (MWCO 12 KDa) suggested a sustained release of the Cur over 72 h in pH 7.4 PBS following the Weibull drug release kinetics. In addition, the ROS generating capabilities upon application of blue light (460 nm) and resulting cytotoxicity were evaluated in MCF-7 cell lines. The Cur-loaded liposome exhibited significant ROS generation and cytotoxicity to the cancer cells than free curcumin. Thus, the Cur-loaded liposomes could be used to treat breast cancer with photodynamic therapy.
Language	English
Publishing date	2022-08-27
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2149918-4
ISSN	1873-1597 ; 1572-1000
ISSN (online)	1873-1597
ISSN	1572-1000
DOI	10.1016/j.pdpdt.2022.103091
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Article ; Online: Estrogen suppresses breast cancer proliferation through GPER / p38 MAPK axis during hypoxia.

Sathya, S / Sudhagar, S / Lakshmi, B S

Molecular and cellular endocrinology

2015 Volume 417, Page(s) 200–210

Abstract: Breast cancer cells frequently experience hypoxia which is associated with resistance to hormonal therapy and poor clinical prognosis, making it important to understand the function of estrogen under hypoxic condition. Here, we demonstrate that estrogen ... ...

Abstract	Breast cancer cells frequently experience hypoxia which is associated with resistance to hormonal therapy and poor clinical prognosis, making it important to understand the function of estrogen under hypoxic condition. Here, we demonstrate that estrogen suppresses breast cancer cell growth under hypoxia, through inhibition at G1/S phase of cell cycle, by elevation of p21 expression. The involvement of GPER in estrogen mediated growth arrest was elucidated using specific ligands and siRNA. Although, estrogen was observed to activate both p44/42 and p38 MAPK signaling, pharmacological inhibition and silencing of p38 MAPK abrogated the induction of p21 expression and growth arrest, during hypoxia. The involvement of estrogen induced ROS in the p38 MAPK mediated p21 expression and cell growth arrest was established by observing that scavenging of ROS by NAC abrogated p38 MAPK activation and p21 expression during hypoxia. In conclusion, Estrogen suppresses breast cancer growth by inhibiting G1/S phase transition through GPER/ROS/p38 MAPK/p21 mediated signaling during hypoxic condition.
MeSH term(s)	Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Cell Cycle/drug effects ; Cell Hypoxia/drug effects ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Cyclin-Dependent Kinase Inhibitor p21/genetics ; Cyclin-Dependent Kinase Inhibitor p21/metabolism ; Estrogens/pharmacology ; Female ; Gene Expression Regulation, Neoplastic/drug effects ; Humans ; MAP Kinase Signaling System/drug effects ; MCF-7 Cells ; Reactive Oxygen Species/metabolism ; Receptors, Estrogen/genetics ; Receptors, Estrogen/metabolism ; Receptors, G-Protein-Coupled/genetics ; Receptors, G-Protein-Coupled/metabolism
Chemical Substances	CDKN1A protein, human ; Cyclin-Dependent Kinase Inhibitor p21 ; Estrogens ; GPER1 protein, human ; Reactive Oxygen Species ; Receptors, Estrogen ; Receptors, G-Protein-Coupled
Language	English
Publishing date	2015-09-30
Publishing country	Ireland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2015.09.032
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Zs.A 1127: Show issues

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Article: Generation of biologically active recombinant human OCT4 protein from E. coli

Dey, Chandrima / Thool, Madhuri / Bhattacharyya, Srirupa / Sudhagar, S / Thummer, Rajkumar P

3 Biotech. 2021 May, v. 11, no. 5

2021

Abstract	Octamer-binding transcription factor 4 (OCT4) is vital for early embryonic development and is a master regulator of pluripotency in embryonic stem cells. Notably, OCT4 is a key reprogramming factor to derive induced pluripotent stem cells, which have tremendous prospects in regenerative medicine. In the current study, we report heterologous expression and purification of human OCT4 in E. coli to produce pure recombinant protein under native conditions. To achieve this, the 1083 bp coding sequence of the human OCT4 gene was codon-optimized for heterologous expression in E. coli. The codon-optimized sequence was fused with fusion tags, namely a cell-penetrating peptide sequence for intracellular delivery, a nuclear localization sequence for intranuclear delivery, and a His-tag for affinity purification. Subsequently, the codon-optimized sequence and the fusion tags were cloned in the protein expression vector, pET28a(+), and transformed into E. coli strain BL21(DE3) for expression. The recombinant OCT4 protein was purified from the soluble fraction under native conditions using immobilized metal ion affinity chromatography in a facile manner, and its identity was confirmed by Western blotting and mass spectrometry. Furthermore, the secondary structure of the recombinant protein was analyzed using far ultraviolet circular dichroism spectroscopy, which confirmed that the purified fusion protein maintained a secondary structure conformation, and it predominantly composed of α-helices. Next, the recombinant OCT4 protein was applied to human cells, and was found that it was able to enter the cells and translocate to the nucleus. Furthermore, the biological activity of the transduced OCT4 protein was also demonstrated on human cells. This recombinant tool can substitute for genetic and viral forms of OCT4 to enable the derivation of integration-free pluripotent cells. It can also be used to elucidate its biological role in various cellular processes and diseases and for structural and biochemical studies.
Keywords	Escherichia coli ; affinity chromatography ; bioactive properties ; circular dichroism spectroscopy ; embryogenesis ; genes ; genetic vectors ; heterologous gene expression ; humans ; mass spectrometry ; medicine ; nuclear localization signals ; recombinant proteins ; transcription factors
Language	English
Dates of publication	2021-05
Size	p. 207.
Publishing place	Springer International Publishing
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2600522-0
ISSN	2190-5738 ; 2190-572X
ISSN (online)	2190-5738
ISSN	2190-572X
DOI	10.1007/s13205-021-02758-z
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