TY - JOUR T1 - Preferential Expansion of Human CD34CD133CD90 Hematopoietic Stem Cells Enhances Gene-Modified Cell Frequency for Gene Therapy. JF - Hum Gene Ther Y1 - 2022 A1 - Christopher, Abisha Crystal A1 - Venkatesan, Vigneshwaran A1 - Karuppusamy, Karthik V A1 - Srinivasan, Saranya A1 - Babu, Prathibha A1 - Azhagiri, Manoj Kumar K A1 - Chambayil, Karthik A1 - Bagchi, Abhirup A1 - Rajendiran, Vignesh A1 - Ravi, Nithin Sam A1 - Kumar, Sanjay A1 - Marepally, Srujan Kumar A1 - Mohankumar, Kumarasamypet Murugesan A1 - Srivastava, Alok A1 - Velayudhan, Shaji R A1 - Thangavel, Saravanabhavan KW - Animals KW - Antigens, CD34 KW - Fetal Blood KW - Genetic Therapy KW - Hematopoietic Stem Cell Transplantation KW - Hematopoietic Stem Cells KW - Humans KW - Mice KW - Mice, Inbred NOD KW - Mice, SCID AB -

CD34CD133CD90 hematopoietic stem cells (HSCs) are responsible for long-term multilineage hematopoiesis, and the high frequency of gene-modified HSCs is crucial for the success of hematopoietic stem and progenitor cell (HSPC) gene therapy. However, the culture and gene manipulation steps of HSPC graft preparation significantly reduce the frequency of HSCs, thus necessitating large doses of HSPCs and reagents for the manipulation. In this study, we identified a combination of small molecules, Resveratrol, UM729, and SR1 that preferentially expands CD34CD133CD90 HSCs over other subpopulations of adult HSPCs in culture. The preferential expansion enriches the HSCs in culture, enhances the adhesion, and results in a sixfold increase in the long-term engraftment in NSG mice. Further, the culture-enriched HSCs are more responsive to gene modification by lentiviral transduction and gene editing, increasing the frequency of gene-modified HSCs up to 10-fold . The yield of gene-modified HSCs obtained by the culture enrichment is similar to the sort-purification of HSCs and superior to Cyclosporin-H treatment. Our study addresses a critical challenge of low frequency of gene modified HSCs in HSPC graft by developing and demonstrating a facile HSPC culture condition that increases the frequency of gene-modified cells . This strategy will improve the outcome of HSPC gene therapy and also simplify the gene manipulation process.

VL - 33 IS - 3-4 ER - TY - JOUR T1 - Pharmacological intervention in young adolescents rescues synaptic physiology and behavioural deficits in Syngap1 mice. JF - Exp Brain Res Y1 - 2021 A1 - Verma, Vijaya A1 - Kumar, M J Vijay A1 - Sharma, Kavita A1 - Rajaram, Sridhar A1 - Muddashetty, Ravi A1 - Manjithaya, Ravi A1 - Behnisch, Thomas A1 - Clement, James P AB -

Haploinsufficiency in SYNGAP1 is implicated in intellectual disability (ID) and autism spectrum disorder (ASD) and affects the maturation of dendritic spines. The abnormal spine development has been suggested to cause a disbalance of excitatory and inhibitory (E/I) neurotransmission at distinct developmental periods. In addition, E/I imbalances in Syngap1 mice might be due to abnormalities in K-Cl co-transporter function (NKCC1, KCC2), in a maner similar to the murine models of Fragile-X and Rett syndromes. To study whether an altered intracellular chloride ion concentration represents an underlying mechanism of modified function of GABAergic synapses in Dentate Gyrus Granule Cells of Syngap1 recordings were performed at different developmental stages of the mice. We observed depolarised neurons at P14-15 as illustrated by decreased Cl reversal potential in Syngap1 mice. The KCC2 expression was decreased compared to Wild-type (WT) mice at P14-15. The GSK-3β inhibitor, 6-bromoindirubin-3'-oxime (6BIO) that crosses the blood-brain barrier, was tested to restore the function of GABAergic synapses. We discovered that the intraperitoneal administration of 6BIO during the critical period or young adolescents [P30 to P80 (4-week to 10-week)] normalised an altered E/I balance, the deficits of synaptic plasticity, and behavioural performance like social novelty, anxiety, and memory of the Syngap1 mice. In summary, altered GABAergic function in Syngap1 mice is due to reduced KCC2 expression leading to an increase in the intracellular chloride concentration that can be counteracted by the 6BIO, which restored cognitive, emotional, and social symptoms by pharmacological intervention, particularly in adulthood.

ER - TY - JOUR T1 - Proteome plasticity in response to persistent environmental change. JF - Mol Cell Y1 - 2021 A1 - Domnauer, Matthew A1 - Zheng, Fan A1 - Li, Liying A1 - Zhang, Yanxiao A1 - Chang, Catherine E A1 - Unruh, Jay R A1 - Conkright-Fincham, Juliana A1 - McCroskey, Scott A1 - Florens, Laurence A1 - Zhang, Ying A1 - Seidel, Christopher A1 - Fong, Benjamin A1 - Schilling, Birgit A1 - Sharma, Rishi A1 - Ramanathan, Arvind A1 - Si, Kausik A1 - Zhou, Chuankai KW - Acclimatization KW - Adaptation, Physiological KW - Animals KW - Environmental Exposure KW - Gene Expression Regulation, Fungal KW - Hot Temperature KW - Proteome KW - Saccharomycetales KW - Stress, Physiological KW - Transcriptome AB -

Temperature is a variable component of the environment, and all organisms must deal with or adapt to temperature change. Acute temperature change activates cellular stress responses, resulting in refolding or removal of damaged proteins. However, how organisms adapt to long-term temperature change remains largely unexplored. Here we report that budding yeast responds to long-term high temperature challenge by switching from chaperone induction to reduction of temperature-sensitive proteins and re-localizing a portion of its proteome. Surprisingly, we also find that many proteins adopt an alternative conformation. Using Fet3p as an example, we find that the temperature-dependent conformational difference is accompanied by distinct thermostability, subcellular localization, and, importantly, cellular functions. We postulate that, in addition to the known mechanisms of adaptation, conformational plasticity allows some polypeptides to acquire new biophysical properties and functions when environmental change endures.

VL - 81 IS - 16 ER - TY - JOUR T1 - Psychiatric symptoms and syndromes transcending diagnostic boundaries in Indian multiplex families: The cohort of ADBS study. JF - Psychiatry Res Y1 - 2021 A1 - Sreeraj, Vanteemar S A1 - Holla, Bharath A1 - Ithal, Dhruva A1 - Nadella, Ravi Kumar A1 - Mahadevan, Jayant A1 - Balachander, Srinivas A1 - Ali, Furkhan A1 - Sheth, Sweta A1 - Narayanaswamy, Janardhanan C A1 - Venkatasubramanian, Ganesan A1 - John, John P A1 - Varghese, Mathew A1 - Benegal, Vivek A1 - Jain, Sanjeev A1 - Reddy, Yc Janardhan A1 - Viswanath, Biju AB -

Syndromes of schizophrenia, bipolar disorder, obsessive-compulsive disorder, substance use disorders and Alzheimer's dementia are highly heritable. About 10-20% of subjects have another affected first degree relative (FDR), and thus represent a 'greater' genetic susceptibility. We screened 3583 families to identify 481 families with multiple affected members, assessed 1406 individuals in person, and collected information systematically about other relatives. Within the selected families, a third of all FDRs were affected with serious mental illness. Although similar diagnoses aggregated within families, 62% of the families also had members with other syndromes. Moreover, 15% of affected individuals met criteria for co-occurrence of two or more syndromes, across their lifetime. Using dimensional assessments, we detected a range of symptom clusters in both affected and unaffected individuals, and across diagnostic categories. Our findings suggest that in multiplex families, there is considerable heterogeneity of clinical syndromes, as well as sub-threshold symptoms. These families would help provide an opportunity for further research using both genetic analyses and biomarkers.

VL - 296 ER - TY - JOUR T1 - A perspective on challenges and opportunities in characterizing oral cancer stem cells. JF - Front Biosci (Landmark Ed) Y1 - 2020 A1 - Sadasivam, Subhashini A1 - Subramanian, Ramaswamy AB -

Cancer stem cells (CSCs) or tumor-initiating cells (TICs) represent a minority population of cells in a tumor that can self-renew and re-create the heterogeneity of the entire tumor. Cell lines, patient-derived tumor cells, and patient-derived xenografts have all been used to isolate presumptive CSC populations from different tumor types. Because of their purported roles in tumor recurrence and prognosis, numerous efforts have centered around reliably identifying CSCs using cell surface markers, and in using genomics tools to identify molecular features unique to these cells. In this brief review, we will discuss different markers, CD44, ALDH1, CD271 and others that have used for identifying and isolating CSCs from primary head & neck and oral squamous cell carcinomas. In particular, we focus on the challenges associated with these experiments as this will be useful to researchers attempting similar isolations. We also discuss some important molecular features gleaned from studying these CSCs such as the expression of stem cell-related markers, loss of cell adhesion and terminal differentiation markers, and the presence of both epithelial and epithelial-to-mesenchymal transition (EMT) features.

VL - 25 ER - TY - JOUR T1 - The primary cilium dampens proliferative signaling and represses a G2/M transcriptional network in quiescent myoblasts. JF - BMC Mol Cell Biol Y1 - 2020 A1 - Venugopal, Nisha A1 - Ghosh, Ananga A1 - Gala, Hardik A1 - Aloysius, Ajoy A1 - Vyas, Neha A1 - Dhawan, Jyotsna AB -

BACKGROUND: Reversible cell cycle arrest (quiescence/G0) is characteristic of adult stem cells and is actively controlled at multiple levels. Quiescent cells also extend a primary cilium, which functions as a signaling hub. Primary cilia have been shown to be important in multiple developmental processes, and are implicated in numerous developmental disorders. Although the association of the cilium with G0 is established, the role of the cilium in the control of the quiescence program is still poorly understood.

RESULTS: Primary cilia are dynamically regulated across different states of cell cycle exit in skeletal muscle myoblasts: quiescent myoblasts elaborate a primary cilium in vivo and in vitro, but terminally differentiated myofibers do not. Myoblasts where ciliogenesis is ablated using RNAi against a key ciliary assembly protein (IFT88) can exit the cell cycle but display an altered quiescence program and impaired self-renewal. Specifically, the G0 transcriptome in IFT88 knockdown cells is aberrantly enriched for G2/M regulators, suggesting a focused repression of this network by the cilium. Cilium-ablated cells also exhibit features of activation including enhanced activity of Wnt and mitogen signaling and elevated protein synthesis via inactivation of the translational repressor 4E-BP1.

CONCLUSIONS: Taken together, our results show that the primary cilium integrates and dampens proliferative signaling, represses translation and G2/M genes, and is integral to the establishment of the quiescence program.

VL - 21 IS - 1 ER - TY - JOUR T1 - Post-transcriptional regulation in planarian stem cells. JF - Semin Cell Dev Biol Y1 - 2019 A1 - Krishna, Srikar A1 - Palakodeti, Dasaradhi A1 - Solana, Jordi AB -

Planarians are known for their immense regenerative abilities. A pluripotent stem cell population provides the cellular source for this process, as well as for the homeostatic cell turnover of the animals. These stem cells, known as neoblasts, present striking similarities at the morphological and molecular level to germ cells, but however, give rise to somatic tissue. Many RNA binding proteins known to be important for germ cell biology are also required for neoblast function, highlighting the importance of post-transcriptional regulation for stem cell control. Many of its aspects, including alternative splicing, alternative polyadenylation, translational control and mRNA deadenylation, as well as small RNAs such as microRNAs and piRNA are critical for stem cells. Their inhibition often abrogates both regeneration and cell turnover, resulting in lethality. Some of aspects of post-transcriptional regulation are conserved from planarian to mammalian stem cells.

VL - 87 ER - TY - JOUR T1 - PAI1 mediates fibroblast-mast cell interactions in skin fibrosis. JF - J Clin Invest Y1 - 2018 A1 - Pincha, Neha A1 - Hajam, Edries Yousaf A1 - Badarinath, Krithika A1 - Batta, Surya Prakash Rao A1 - Masudi, Tafheem A1 - Dey, Rakesh A1 - Andreasen, Peter A1 - Kawakami, Toshiaki A1 - Samuel, Rekha A1 - George, Renu A1 - Danda, Debashish A1 - Jacob, Paul Mazhuvanchary A1 - Jamora, Colin AB -

Fibrosis is a prevalent pathological condition arising from the chronic activation of fibroblasts. This activation results from the extensive intercellular crosstalk mediated by both soluble factors and direct cell-cell connections. Prominent among these are the interactions of fibroblasts with immune cells, in which the fibroblast-mast cell connection, although acknowledged, is relatively unexplored. We have used a Tg mouse model of skin fibrosis, based on expression of the transcription factor Snail in the epidermis, to probe the mechanisms regulating mast cell activity and the contribution of these cells to this pathology. We have discovered that Snail-expressing keratinocytes secrete plasminogen activator inhibitor type 1 (PAI1), which functions as a chemotactic factor to increase mast cell infiltration into the skin. Moreover, we have determined that PAI1 upregulates intercellular adhesion molecule type 1 (ICAM1) expression on dermal fibroblasts, rendering them competent to bind to mast cells. This heterotypic cell-cell adhesion, also observed in the skin fibrotic disorder scleroderma, culminates in the reciprocal activation of both mast cells and fibroblasts, leading to the cascade of events that promote fibrogenesis. Thus, we have identified roles for PAI1 in the multifactorial program of fibrogenesis that expand its functional repertoire beyond its canonical role in plasmin-dependent processes.

VL - 128 IS - 5 ER - TY - CHAP T1 - Prevention of Metal Exposure: Chelating Agents and Barrier Creams T2 - Metal Allergy: From Dermatitis to Implant and Device Failure Y1 - 2018 A1 - Mahato, Manohar A1 - Sherman, Nicholas E. A1 - Kiran Kumar Mudnakudu, N. A1 - Joshi, Nitin A1 - Briand, Elisabeth A1 - Karp, Jeffrey M. A1 - Vemula, Praveen Kumar ED - Chen, Jennifer K ED - Thyssen, Jacob P. AB -

Metals are a group of elements which are ubiquitous in modern life. They are used in the fields of cosmetics, water purification, medicine, paint, food products, pesticides, and almost innumerable others. As the use of metals has increased in recent decades, so has human exposure to these elements. Metals such as mercury, lead, arsenic, nickel, and others have been implicated in negatively affecting human homeostasis by causing chronic inflammatory diseases, among other serious conditions. Both acute and chronic metal toxicity in vital organs could arise from local or systemic exposure to numerous metals. Although some metals have health benefits, overaccumulation of metals in body tissues can result in deleterious, toxic effects. Most exposure to metals occurs via cutaneous, inhalation, or oral routes. At the highest risk of negative effects of exposure are pregnant women and children. To ameliorate or prevent the toxic effects of metals, chelating agents and barrier creams are used widely in medical practice today. In this chapter, we will discuss preventing metal toxicity from overexposure via chelation therapy and skin barrier creams.

JF - Metal Allergy: From Dermatitis to Implant and Device Failure PB - Springer International Publishing CY - Cham SN - 978-3-319-58503-1 UR - https://doi.org/10.1007/978-3-319-58503-1_18 ER - TY - JOUR T1 - Prevention of pesticide-induced neuronal dysfunction and mortality with nucleophilic poly-Oxime topical gel. JF - Sci Adv Y1 - 2018 A1 - Thorat, Ketan A1 - Pandey, Subhashini A1 - Chandrashekharappa, Sandeep A1 - Vavilthota, Nikitha A1 - Hiwale, Ankita A A1 - Shah, Purna A1 - Sreekumar, Sneha A1 - Upadhyay, Shubhangi A1 - Phuntsok, Tenzin A1 - Mahato, Manohar A1 - Mudnakudu-Nagaraju, Kiran K A1 - Sunnapu, Omprakash A1 - Vemula, Praveen K AB -

Organophosphate-based pesticides inhibit acetylcholinesterase (AChE), which plays a pivotal role in neuromuscular function. While spraying in the field, farmworkers get exposed to pesticides through the dermal route. Internalized pesticide inhibits AChE, which leads to neurotoxicity, cardiotoxicity, cognitive dysfunction, loss of endurance, and death in severe cases. Here, we present a nucleophilic pyridine-2-aldoxime-functionalized chitosan-based topical gel (-Oxime gel) that rapidly deactivates organophosphates, methyl parathion (MPT), on the skin of rats, which leads to reduced AChE inhibition in the blood and tissues. Testing the robustness of -Oxime gel, we report reduction in AChE inhibition following repeated dermal administration of MPT in the presence of -Oxime gel. Furthermore, -Oxime gel prevented MPT-induced neuromuscular dysfunction, loss of endurance, and locomotor coordination. We observe a 100% survival in rats following topical MPT administration in the presence of -Oxime gel. This prophylactic gel may therefore help farmworkers by limiting pesticide-induced toxicity and mortality.

VL - 4 IS - 10 ER -