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Article ; Online: Thermal Shift Assay in Ferroptosis.

Bammidi, Sridhar / Hose, Stacey / Handa, James T / Sinha, Debasish / Ghosh, Sayan

Methods in molecular biology (Clifton, N.J.)

2023 Volume 2712, Page(s) 179–186

Abstract: Ferroptosis is a recently described process of cell death that is dependent on unregulated cellular iron accumulation with induction of oxidative stress. Ferroptosis has been linked to several human diseases; therefore, investigations aimed at better ... ...

Abstract	Ferroptosis is a recently described process of cell death that is dependent on unregulated cellular iron accumulation with induction of oxidative stress. Ferroptosis has been linked to several human diseases; therefore, investigations aimed at better understanding the pathway and elucidating avenues for future drug development are warranted. Current assays that target ferroptosis/oxidative stress in cells is limited to western blotting and imaging techniques, and unfortunately provide only a broad understanding that is insufficient to effectively assess novel drugs (ligands). Specifically, these assays do not provide insights about ligand interactions with specific proteins associated with these processes. Herein, we discuss a cell-based thermal shift assay that enables screening of ligands under specific cellular conditions for targeting ferroptosis and/or oxidative stress pathways. These data would provide detailed preliminary evidence required for drug development that targets this pathway.
MeSH term(s)	Humans ; Ferroptosis ; Biological Assay ; Blotting, Western ; Cell Death ; Drug Development
Language	English
Publishing date	2023-08-04
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3433-2_16
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Article ; Online: Transplantation Efficacy of Human Ciliary Epithelium Cells from Fetal Eye and Lin-ve Stem Cells from Umbilical Cord Blood in the Murine Retinal Degeneration Model of Laser Injury.

Bammidi, Sridhar / Bali, Parul / Kalra, Jaswinder / Anand, Akshay

Cell transplantation

2020 Volume 29, Page(s) 963689720946031

Abstract: A number of degenerative conditions affecting the neural retina including age-related macular degeneration have no successful treatment, resulting in partial or complete vision loss. There are a number of stem cell replacement strategies for recovery of ... ...

Abstract	A number of degenerative conditions affecting the neural retina including age-related macular degeneration have no successful treatment, resulting in partial or complete vision loss. There are a number of stem cell replacement strategies for recovery of retinal damage using cells from variable sources. However, literature is still deficit in the comparison of efficacy of types of stem cells. The purpose of the study was to compare the therapeutic efficacy of undifferentiated cells, i.e., lineage negative stem cells (Lin-ve SC) with differentiated neurosphere derived from ciliary epithelium (CE) cells on retinal markers associated with laser-induced retinal injury. Laser-induced photocoagulation was carried out to disrupt Bruch's membrane and retinal pigmented epithelium in C57BL/6 mouse model. Lineage negative cells were isolated from human umbilical cord blood, whereas neurospheres were derived from CE of post-aborted human eyeballs. The cells were then transplanted into subretinal space to study their effect on injury. Markers of neurotropic factors, retina, apoptosis, and proliferation were analyzed after injury and transplantation. mRNA expression was also analyzed by real-time polymerase chain reaction at 1 week, and 3-month immunohistochemistry was evaluated at 1-week time point. CE cell transplantation showed enhanced differentiation of rods and retinal glial cells. However, Lin-ve cells exerted paracrine-dependent modulation of neurotrophic factors, which is possibly mediated by antiapoptotic and proliferative effects. In conclusion, CE transplantation showed superior regenerative outcome in comparison to Lin-ve SC for rescue of artificially injured rodent retinal cells. It is imperative that this source for transplantation may be extensively studied in various doses and additional retinal degeneration models for prospective clinical applications.
MeSH term(s)	Animals ; Apoptosis ; Cell Differentiation ; Cell Lineage ; Cell Proliferation ; Cilia/metabolism ; Ciliary Neurotrophic Factor/metabolism ; Disease Models, Animal ; Epithelial Cells/cytology ; Epithelial Cells/transplantation ; Eye/embryology ; Fetal Blood/cytology ; Fetus/embryology ; Humans ; Lasers/adverse effects ; Male ; Mice, Inbred C57BL ; Nerve Growth Factors/metabolism ; Retinal Degeneration/pathology ; Retinal Degeneration/therapy ; Retinal Ganglion Cells/pathology ; Spheroids, Cellular/cytology ; Stem Cell Transplantation ; Stem Cells/cytology
Chemical Substances	Ciliary Neurotrophic Factor ; Nerve Growth Factors
Language	English
Publishing date	2020-10-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1135816-6
ISSN	1555-3892 ; 0963-6897
ISSN (online)	1555-3892
ISSN	0963-6897
DOI	10.1177/0963689720946031
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Article ; Online: A CD33 Antigen-Targeted AAV6 Vector Expressing an Inducible Caspase-9 Suicide Gene Is Therapeutic in a Xenotransplantation Model of Acute Myeloid Leukemia.

Khan, Nusrat / Bammidi, Sridhar / Jayandharan, Giridhara R

Bioconjugate chemistry

2019 Volume 30, Issue 9, Page(s) 2404–2416

Abstract: Current chemotherapeutic regimens for acute myeloid leukemia (AML) have been modestly effective in patients and are associated with poor long-term survival (<30% at 5 years). Viral vector-based suicide gene therapy is an attractive option, if these ... ...

Abstract	Current chemotherapeutic regimens for acute myeloid leukemia (AML) have been modestly effective in patients and are associated with poor long-term survival (<30% at 5 years). Viral vector-based suicide gene therapy is an attractive option, if these vectors can target the AML cells with high specificity and efficiency. In this study, we have developed a receptor-specific adeno-associated virus (AAV) based vector to target the CD33 antigen which is overexpressed in leukemic cells. A targeting peptide was rationally designed from the antigen-binding regions of a CD33 monoclonal antibody. This peptide was further expressed on the capsid of the AAV6 vector, since this serotype was most efficient among AAV1-rh10 vectors to infect the pro-monocytic, human myeloid leukemia cells (U937). AAV6-CD33 vectors expressing a suicide gene, the inducible caspase 9 (iCasp9), and its prodrug AP20187 significantly reduced (∼59%) the viability of U937 cells. To further test its efficacy and specificity
MeSH term(s)	Animals ; Caspase 9/genetics ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Dependovirus/genetics ; Gene Expression ; Genes, Transgenic, Suicide/genetics ; Genetic Therapy/methods ; Genetic Vectors/genetics ; Humans ; Leukemia, Myeloid, Acute/genetics ; Leukemia, Myeloid, Acute/pathology ; Leukemia, Myeloid, Acute/therapy ; Sialic Acid Binding Ig-like Lectin 3/genetics ; Zebrafish
Chemical Substances	Sialic Acid Binding Ig-like Lectin 3 ; Caspase 9 (EC 3.4.22.-)
Language	English
Publishing date	2019-09-04
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1024041-x
ISSN	1520-4812 ; 1043-1802
ISSN (online)	1520-4812
ISSN	1043-1802
DOI	10.1021/acs.bioconjchem.9b00511
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Article ; Online: Human Fetal Pigmented Ciliary Epithelium Stem Cells have Regenerative Capacity in the Murine Retinal Degeneration Model of Laser Injury.

Bammidi, Sridhar / Modgil, Shweta / Kalra, Jaswinder / Anand, Akshay

Current neurovascular research

2019 Volume 16, Issue 3, Page(s) 187–193

Abstract: Background: Retinal degeneration and related eye disorders have limited treatment interventions. Since stem cell therapy has shown promising results, ciliary epithelium (CE) derived stem cells could be a better choice given the fact that cells from eye ... ...

Abstract	Background: Retinal degeneration and related eye disorders have limited treatment interventions. Since stem cell therapy has shown promising results, ciliary epithelium (CE) derived stem cells could be a better choice given the fact that cells from eye niche can better integrate with the degenerating retina, rewiring the synaptic damage. Objective: To test the effect of human fetal pigmented ciliary epithelium-derived neurospheres in the mouse model of laser-induced retinal degeneration. Methods: C57 male mice were subjected to retinal injury by Laser photocoagulation. Human fetal pigmented ciliary epithelium was obtained from post-aborted human eyeballs and cultured with epidermal growth factor (rhEGF) and fibroblast growth factor (rhFGF). The six day neurospheres were isolated, dissociated and transplanted into the subretinal space of the laser injured mice at the closest proximity to Laser shots. Mice were analyzed for functional vision through electroretinogram (ERG) and sacrificed at 1 week and 12 week time points. Retinal, Neurotropic, Apoptotic and proliferation markers were analysed using real-time polymerase chain reaction (PCR). Results: The CE neurospheres showed an increase in the expression of candidate genes analyzed in the study at 1 week time point, which sustained for longer time point of 12 weeks. Conclusion: We showed the efficacy of human CE cells in the regeneration of retinal degeneration in murine model for the first time. CE cells need to be explored comprehensively both in disease and degeneration.
MeSH term(s)	Animals ; Cells, Cultured ; Cilia/physiology ; Cilia/transplantation ; Epithelial Cells/physiology ; Epithelial Cells/transplantation ; Fetal Stem Cells/chemistry ; Fetal Stem Cells/physiology ; Fetal Stem Cells/transplantation ; Humans ; Lasers/adverse effects ; Male ; Mice ; Mice, Inbred C57BL ; Nerve Regeneration/physiology ; Retinal Degeneration/etiology ; Retinal Degeneration/pathology ; Retinal Degeneration/therapy ; Retinal Pigment Epithelium/physiology ; Retinal Pigment Epithelium/transplantation ; Stem Cell Transplantation/methods
Language	English
Publishing date	2019-07-01
Publishing country	United Arab Emirates
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2296350-9
ISSN	1875-5739 ; 1567-2026
ISSN (online)	1875-5739
ISSN	1567-2026
DOI	10.2174/1567202616666190618123931
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Zs.A 6646: Show issues

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Article: AAV6 Vexosomes Mediate Robust Suicide Gene Delivery in a Murine Model of Hepatocellular Carcinoma.

Khan, Nusrat / Maurya, Shubham / Bammidi, Sridhar / Jayandharan, Giridhara R

Molecular therapy. Methods & clinical development

2020 Volume 17, Page(s) 497–504

Abstract: During recombinant Adeno-associated virus (AAV) production, a proportionately large amount of vectors is released in the culture supernatant, which is often discarded. It has been shown that these vectors often associate with vesiculated structures, such ...

Abstract	During recombinant Adeno-associated virus (AAV) production, a proportionately large amount of vectors is released in the culture supernatant, which is often discarded. It has been shown that these vectors often associate with vesiculated structures, such as exosomes. Exosome-associated AAV (vexosomes) represent an additional gene-delivery platform. The efficiency of such vexosomes in suicide gene therapy is unexplored. In the present study, we have generated AAV serotype 6 vexosomes containing an inducible caspase 9 (
Language	English
Publishing date	2020-03-13
Publishing country	United States
Document type	Journal Article
ZDB-ID	2872938-9
ISSN	2329-0501 ; 2329-0501
ISSN (online)	2329-0501
ISSN	2329-0501
DOI	10.1016/j.omtm.2020.03.006
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Zs.A 7107: Show issues

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Article ; Online: Combination Suicide Gene Delivery with an Adeno-Associated Virus Vector Encoding Inducible Caspase-9 and a Chemical Inducer of Dimerization Is Effective in a Xenotransplantation Model of Hepatocellular Carcinoma.

Khan, Nusrat / Bammidi, Sridhar / Chattopadhyay, Sourav / Jayandharan, Giridhara R

Bioconjugate chemistry

2019 Volume 30, Issue 6, Page(s) 1754–1762

Abstract: Current treatment approaches for hepatocellular carcinoma (HCC) have a narrow therapeutic index and alternate modes of treatment are thus required. We have utilized a gene delivery vector containing inducible caspase 9 (iCasp9) gene, which is a synthetic ...

Abstract	Current treatment approaches for hepatocellular carcinoma (HCC) have a narrow therapeutic index and alternate modes of treatment are thus required. We have utilized a gene delivery vector containing inducible caspase 9 (iCasp9) gene, which is a synthetic analogue based on the mammalian caspase 9 and fused to a human FK506 binding protein that allows its conditional dimerization to a synthetic, small molecule [chemical inducer of dimerization, AP20187] and results in target cell apoptosis. In our studies, we have tested these synthetic vectors based on an adeno-associated virus platform for their potential anti-tumorigenic effect in human HCC cells in vitro and in a HCC tumor model developed in nude mice. Our data demonstrates that the iCasp9-AP20187 bioconjugate is able to trigger terminal effectors of cellular apoptosis and presents a viable approach for the potential treatment of HCC.
MeSH term(s)	Animals ; Carcinoma, Hepatocellular/genetics ; Carcinoma, Hepatocellular/therapy ; Caspase 9/genetics ; Cell Line, Tumor ; Dependovirus/genetics ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors/genetics ; Genetic Vectors/therapeutic use ; Humans ; Liver Neoplasms/genetics ; Liver Neoplasms/therapy ; Mice, Inbred BALB C ; Mice, Nude ; Tacrolimus Binding Proteins/genetics
Chemical Substances	Caspase 9 (EC 3.4.22.-) ; Tacrolimus Binding Proteins (EC 5.2.1.-)
Language	English
Publishing date	2019-06-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1024041-x
ISSN	1520-4812 ; 1043-1802
ISSN (online)	1520-4812
ISSN	1043-1802
DOI	10.1021/acs.bioconjchem.9b00291
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Zs.A 3400: Show issues

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Article ; Online: MicroRNA-based recombinant AAV vector assembly improves efficiency of suicide gene transfer in a murine model of lymphoma.

Khan, Nusrat / Cheemadan, Sabna / Saxena, Himanshi / Bammidi, Sridhar / Jayandharan, Giridhara R

Cancer medicine

2020 Volume 9, Issue 9, Page(s) 3188–3201

Abstract: Recent success in clinical trials with recombinant Adeno-associated virus (AAV)-based gene therapy has redirected efforts in optimizing AAV assembly and production, to improve its potency. We reasoned that inclusion of a small RNA during vector assembly, ...

Abstract	Recent success in clinical trials with recombinant Adeno-associated virus (AAV)-based gene therapy has redirected efforts in optimizing AAV assembly and production, to improve its potency. We reasoned that inclusion of a small RNA during vector assembly, which specifically alters the phosphorylation status of the packaging cells may be beneficial. We thus employed microRNAs (miR-431, miR-636) identified by their ability to bind AAV genome and also dysregulate Mitogen-activated protein kinase (MAPK) signaling during vector production, by a global transcriptome study in producer cells. A modified vector assembly protocol incorporating a plasmid encoding these microRNAs was developed. AAV2 vectors packaged in the presence of microRNA demonstrated an improved gene transfer potency by 3.7-fold, in vitro. Furthermore, AAV6 serotype vectors encoding an inducible caspase 9 suicide gene, packaged in the presence of miR-636, showed a significant tumor regression (~2.2-fold, P < .01) in a syngeneic murine model of T-cell lymphoma. Taken together, we have demonstrated a simple but effective microRNA-based approach to improve the assembly and potency of suicide gene therapy with AAV vectors.
MeSH term(s)	Caspase 9/genetics ; Dependovirus/genetics ; Gene Transfer Techniques ; Genes, Transgenic, Suicide ; Genetic Therapy ; Genetic Vectors/administration & dosage ; Genetic Vectors/genetics ; Humans ; Lymphoma/genetics ; Lymphoma/therapy ; MicroRNAs/genetics ; Transduction, Genetic ; Tumor Cells, Cultured
Chemical Substances	MicroRNAs ; CASP9 protein, human (EC 3.4.22.-) ; Caspase 9 (EC 3.4.22.-)
Language	English
Publishing date	2020-02-28
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2659751-2
ISSN	2045-7634 ; 2045-7634
ISSN (online)	2045-7634
ISSN	2045-7634
DOI	10.1002/cam4.2935
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Article: Medical education and training: implications for india.

Anand, Akshay / Bammidi, Sridhar

Annals of neurosciences

2014 Volume 20, Issue 4, Page(s) 133

Language	English
Publishing date	2014-09-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	2576191-2
ISSN	0976-3260 ; 0972-7531
ISSN (online)	0976-3260
ISSN	0972-7531
DOI	10.5214/ans.0972.7531.200402
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Article: Medical education and training: implications for india.

Anand, Akshay / Bammidi, Sridhar

Annals of neurosciences

2014 Volume 21, Issue 1, Page(s) 3–4

Language	English
Publishing date	2014-09-09
Publishing country	United States
Document type	Journal Article
ZDB-ID	2576191-2
ISSN	0976-3260 ; 0972-7531
ISSN (online)	0976-3260
ISSN	0972-7531
DOI	10.5214/ans.0972.7531.210102
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Article: CD34 and CD117 Stemness of Lineage-Negative Cells Reverses Memory Loss Induced by Amyloid Beta in Mouse Model.

Bali, Parul / Bammidi, Sridhar / Banik, Avijit / Nehru, Bimla / Anand, Akshay

Frontiers in behavioral neuroscience

2018 Volume 12, Page(s) 222

Abstract: A majority of the neurodegenerative disorders including Alzheimer's disease are untreatable and occur primarily due to aging and rapidly changing lifestyles. The rodent Alzheimer's disease models are critical for investigating the underlying disease ... ...

Abstract	A majority of the neurodegenerative disorders including Alzheimer's disease are untreatable and occur primarily due to aging and rapidly changing lifestyles. The rodent Alzheimer's disease models are critical for investigating the underlying disease pathology and screening of novel therapeutic targets in preclinical settings. We aimed to characterize the stemness properties of human umbilical cord blood (hUCB) derived lineage-negative (Lin-) stem cells based on CD34 and CD117 expression as well as surface morphology using flow cytometry and scanning electron microscopy, respectively. The efficacy of the stem cells was tested by its capacity to rescue the injury caused by intrahippocampal delivery of varying doses of amyloid beta. The hUCB Lin- stem cells reversed memory loss due to Aβ42-induced injury more effectively at micromolar concentration, and not picomolar concentration. More studies are required to delineate the underlying molecular events associated with hUCB Lin- stem cells.
Language	English
Publishing date	2018-11-01
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2452960-6
ISSN	1662-5153
ISSN	1662-5153
DOI	10.3389/fnbeh.2018.00222
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