Preparation, characterization and cell response studies on bioconjugated 3D protein hydrogels with wide-range stiffness: an approach on cell therapy and cell storage

The success of gene and cell therapy in clinic during the past two decades as well as our expanding ability to manipulate these biomaterials are leading to new therapeutic options for a wide range of inherited and acquired diseases. Combining conventional therapies with this emerging field is a promising strategy to treat those previously-thought untreatable diseases. Traditional Chinese medicine (TCM) has evolved for thousands of years in China and still plays an important role in human health. As part of the active ingredients of TCM, proteins and peptides have attracted long-term enthusiasm of researchers. More recently, they have been utilized in gene and cell therapy, resulting in promising novel strategies to treat both cancer and non-cancer diseases. This manuscript presents a critical review on this field, accompanied with perspectives on the challenges and new directions for future research in this emerging frontier.

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'Off-the-shelf’ allogeneic antigen-specific adoptive T-cell therapy for the treatment of multiple EBV-associated malignancies

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-001608 ◽

2021 ◽

Vol 9 (2) ◽

pp. e001608

Author(s):

Debottam Sinha ◽

Sriganesh Srihari ◽

Kirrliee Beckett ◽

Laetitia Le Texier ◽

Matthew Solomon ◽

...

Keyword(s):

T Cell ◽

Cell Therapy ◽

Adoptive T Cell Therapy ◽

Nuclear Antigen ◽

T Cell Therapy ◽

In Vivo Models ◽

Hla Alleles ◽

Cell Therapies ◽

Wide Range

BackgroundEpstein-Barr virus (EBV), an oncogenic human gammaherpesvirus, is associated with a wide range of human malignancies of epithelial and B-cell origin. Recent studies have demonstrated promising safety and clinical efficacy of allogeneic ‘off-the-shelf’ virus-specific T-cell therapies for post-transplant viral complications.MethodsTaking a clue from these studies, we developed a highly efficient EBV-specific T-cell expansion process using a replication-deficient AdE1-LMPpoly vector that specifically targets EBV-encoded nuclear antigen 1 (EBNA1) and latent membrane proteins 1 and 2 (LMP1 and LMP2), expressed in latency II malignancies.ResultsThese allogeneic EBV-specific T cells efficiently recognized human leukocyte antigen (HLA)-matched EBNA1-expressing and/or LMP1 and LMP2-expressing malignant cells and demonstrated therapeutic potential in a number of in vivo models, including EBV lymphomas that emerged spontaneously in humanized mice following EBV infection. Interestingly, we were able to override resistance to T-cell therapy in vivo using a ‘restriction-switching’ approach, through sequential infusion of two different allogeneic T-cell therapies restricted through different HLA alleles. Furthermore, we have shown that inhibition of the programmed cell death protein-1/programmed death-ligand 1 axis in combination with EBV-specific T-cell therapy significantly improved overall survival of tumor-bearing mice when compared with monotherapy.ConclusionThese findings suggest that restriction switching by sequential infusion of allogeneic T-cell therapies that target EBV through distinct HLA alleles may improve clinical response.

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Electrospun Fiber Scaffolds for Engineering Glial Cell Behavior to Promote Neural Regeneration

Bioengineering ◽

10.3390/bioengineering8010004 ◽

2020 ◽

Vol 8 (1) ◽

pp. 4

Author(s):

Devan L. Puhl ◽

Jessica L. Funnell ◽

Derek W. Nelson ◽

Manoj K. Gottipati ◽

Ryan J. Gilbert

Keyword(s):

Nervous System ◽

Glial Cell ◽

Cell Response ◽

Electrospun Fiber ◽

Neural Regeneration ◽

Cell Phenotype ◽

Comprehensive Overview ◽

Fiber Alignment ◽

Wide Range ◽

Fiber Scaffolds

Electrospinning is a fabrication technique used to produce nano- or micro- diameter fibers to generate biocompatible, biodegradable scaffolds for tissue engineering applications. Electrospun fiber scaffolds are advantageous for neural regeneration because they mimic the structure of the nervous system extracellular matrix and provide contact guidance for regenerating axons. Glia are non-neuronal regulatory cells that maintain homeostasis in the healthy nervous system and regulate regeneration in the injured nervous system. Electrospun fiber scaffolds offer a wide range of characteristics, such as fiber alignment, diameter, surface nanotopography, and surface chemistry that can be engineered to achieve a desired glial cell response to injury. Further, electrospun fibers can be loaded with drugs, nucleic acids, or proteins to provide the local, sustained release of such therapeutics to alter glial cell phenotype to better support regeneration. This review provides the first comprehensive overview of how electrospun fiber alignment, diameter, surface nanotopography, surface functionalization, and therapeutic delivery affect Schwann cells in the peripheral nervous system and astrocytes, oligodendrocytes, and microglia in the central nervous system both in vitro and in vivo. The information presented can be used to design and optimize electrospun fiber scaffolds to target glial cell response to mitigate nervous system injury and improve regeneration.

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Cell rounding causes genomic instability by dissociation of single-stranded DNA-binding proteins

10.1101/463653 ◽

2018 ◽

Author(s):

Qian Guo ◽

Xianglu Liao ◽

Xingwu Wang ◽

Ling Liu ◽

Bao Song

Keyword(s):

Stem Cell ◽

Dna Binding ◽

Cell Therapy ◽

Genomic Instability ◽

Stem Cell Therapy ◽

Binding Proteins ◽

Dna Binding Proteins ◽

Single Stranded Dna ◽

Cell Rounding ◽

Wide Range

AbstractGenomic instability can cause a wide range of diseases, including cancer and cellular senescence, which is also a major challenge in stem cell therapy. However, how a single event can cause extremely high levels of genomic instability remains unclear. Using our developed method, cell in situ electrophoresis (CISE), and models of normal, cancer, and embryonic stem cells, we found that cell rounding as a catastrophic source event ubiquitously observed in vivo and in vitro might lead to large-scale DNA deprotection, genomic instability, chromosomal shattering, cell heterogeneity, and senescent crisis by dissociation of single-stranded DNA-binding proteins (SSBs). Understanding the mechanism may facilitate the development of clinical strategies for cancer therapy, improve the safety of stem cell therapy, and prevent pathological aging.

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Toward Personalized Cell Therapies by Using Stem Cells: Seven Relevant Topics for Safety and Success in Stem Cell Therapy

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/758102 ◽

2012 ◽

Vol 2012 ◽

pp. 1-12 ◽

Cited By ~ 8

Author(s):

Fernando de Sá Silva ◽

Paula Nascimento Almeida ◽

João Vitor Paes Rettore ◽

Claudinéia Pereira Maranduba ◽

Camila Maurmann de Souza ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Therapy ◽

Cell Biology ◽

Stem Cell Biology ◽

Controversial Issues ◽

Immunosuppressive Activity ◽

Cell Therapies ◽

Tumorigenic Potential ◽

Wide Range

Stem cells, both embryonic and adult, due to the potential for application in tissue regeneration have been the target of interest to the world scientific community. In fact, stem cells can be considered revolutionary in the field of medicine, especially in the treatment of a wide range of human diseases. However, caution is needed in the clinical application of such cells and this is an issue that demands more studies. This paper will discuss some controversial issues of importance for achieving cell therapy safety and success. Particularly, the following aspects of stem cell biology will be presented: methods for stem cells culture, teratogenic or tumorigenic potential, cellular dose, proliferation, senescence, karyotyping, and immunosuppressive activity.

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Chemical unfolding of protein domains induces shape change in programmed protein hydrogels

Nature Communications ◽

10.1038/s41467-019-13312-0 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 10

Author(s):

Luai R. Khoury ◽

Ionel Popa

Keyword(s):

Protein Unfolding ◽

Shape Change ◽

Fold Increase ◽

Protein Mechanics ◽

Protein Polymer ◽

Wide Range ◽

Biomechanical Response ◽

Polymer Interactions ◽

Smart Biomaterials ◽

Protein Hydrogels

AbstractProgrammable behavior combined with tailored stiffness and tunable biomechanical response are key requirements for developing successful materials. However, these properties are still an elusive goal for protein-based biomaterials. Here, we use protein-polymer interactions to manipulate the stiffness of protein-based hydrogels made from bovine serum albumin (BSA) by using polyelectrolytes such as polyethyleneimine (PEI) and poly-L-lysine (PLL) at various concentrations. This approach confers protein-hydrogels with tunable wide-range stiffness, from ~10–64 kPa, without affecting the protein mechanics and nanostructure. We use the 6-fold increase in stiffness induced by PEI to program BSA hydrogels in various shapes. By utilizing the characteristic protein unfolding we can induce reversible shape-memory behavior of these composite materials using chemical denaturing solutions. The approach demonstrated here, based on protein engineering and polymer reinforcing, may enable the development and investigation of smart biomaterials and extend protein hydrogel capabilities beyond their conventional applications.

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Stem Cell Therapy in Treating Type 1 Diabetes Mellitus –Potential and Ethical Issues.

Malaysian Journal of Science Health & Technology ◽

10.33102/mjosht.v6i.122 ◽

2021 ◽

Author(s):

Nursuaidah Abdullah ◽

Marjanu Hikmah Elias

Keyword(s):

Stem Cells ◽

Type 1 Diabetes ◽

Stem Cell ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Cell Transplant ◽

Exogenous Insulin ◽

Hematopoietic Stem ◽

Wide Range

Type 1 diabetes (T1D) is a deficiency in insulin production which is mainly due to loss of ?-cell pancreatic islets. Patients with T1D need to be given exogenous insulin regularly. While improvements in the delivery of insulin and glucose monitoring methods have been effective in improving patient safety, insulin therapy is not a cure and is often associated with complications and debilitating hypoglycaemic episodes. Meanwhile, pancreas or islet transplantation as a gold standard only promises temporary freedom from exogenous insulin and suffers from issues of its own. Stem cell therapy may provide a more permanent solution, given stem cells’ immunomodulatory characteristics and ability to self-renew and distinguish into specific cells. In this sense, the therapeutic potentials of stem cells are addressed in this study. These stem cells cover a wide range of treatments for T1D including embryonic stem cells, induced pluripotent stem cells, bone-marrow derived hematopoietic stem cells and multipotent mesenchymal stromal cells. The challenges faced by the current stem cell transplant in T1D treatment and Islamic viewpoints regarding ethics in stem cell research and therapy are also discussed. In conclusion, stem cell therapy offers a safe and efficient alternative treatment for T1D. However, besides the fatwa from Fatwa Committee of Selangor, the lack of Malaysian stem cells ethics should be further addressed.

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A non-structural pure enzyme protein forms a LCST type of stimuli-responsive and reversible hydrogel with novel structure and catalytic activity

10.1101/2021.02.07.430034 ◽

2021 ◽

Author(s):

J. Nie ◽

X. Zhang ◽

Y. Liu ◽

M.A. Schroer ◽

W. Wang ◽

...

Keyword(s):

Catalytic Activity ◽

Electrostatic Interactions ◽

Molecular Engineering ◽

Solution Temperature ◽

Stimuli Responsive ◽

Electron Microscopic ◽

Saxs Data ◽

Wide Range ◽

Protein Hydrogels ◽

Terminal Domains

AbstractHydrogels have a wide range of applications such as in biomedicine, cosmetics and soft electronics. Compared to polymer hydrogels based on covalent bonding, protein hydrogels offer distinct advantages owing to their biocompatibility and better access to molecular engineering. However, pure and natural protein hydrogels have been seldom reported except for structural proteins like collagen and silk fibrin. Here, we report the unusual ability and mechanism of a unique natural enzyme, lipoate-protein ligase A (LplA) of E. coli to self-assemble into a stimuli-responsive and reversible hydrogel of the low critical solution temperature (LCST) type. This is the first globular and catalytic protein found to form a hydrogel in response to temperature, pH and the presence of ions. Protein structure based analysis reveals the key residues responsible for the gel formation and mutational studies confirms the essential roles of hydrogen bonding between the C-terminal domains and electrostatic interactions in the N-terminal domains. Characterization of phase transitions of wild type LplA and its mutants using small angle X-ray scattering (SAXS) yields details of the gelation process from initial dimer formation over a pre-gel-state to full network development. Further electron microscopic analyses and modeling of SAXS data suggest an unusual interlinked ladder-like structure of the macroscopic crosslinking network with dimers as ladder steps. The unique features of this first reported protein hydrogel may open up hitherto inaccessible applications, especially those taking advantage of the inherent catalytic activity of LplA.

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Targeting Programmed Cell Death to Improve Stem Cell Therapy: Implications for Treating Diabetes and Diabetes-Related Diseases

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.809656 ◽

2021 ◽

Vol 9 ◽

Author(s):

Qi Zhang ◽

Xin-xing Wan ◽

Xi-min Hu ◽

Wen-juan Zhao ◽

Xiao-xia Ban ◽

...

Keyword(s):

Stem Cells ◽

Cell Death ◽

Stem Cell ◽

Programmed Cell Death ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Therapeutic Effects ◽

Stem Cell Therapies ◽

Cell Therapies ◽

Wide Range

Stem cell therapies have shown promising therapeutic effects in restoring damaged tissue and promoting functional repair in a wide range of human diseases. Generations of insulin-producing cells and pancreatic progenitors from stem cells are potential therapeutic methods for treating diabetes and diabetes-related diseases. However, accumulated evidence has demonstrated that multiple types of programmed cell death (PCD) existed in stem cells post-transplantation and compromise their therapeutic efficiency, including apoptosis, autophagy, necroptosis, pyroptosis, and ferroptosis. Understanding the molecular mechanisms in PCD during stem cell transplantation and targeting cell death signaling pathways are vital to successful stem cell therapies. In this review, we highlight the research advances in PCD mechanisms that guide the development of multiple strategies to prevent the loss of stem cells and discuss promising implications for improving stem cell therapy in diabetes and diabetes-related diseases.

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Stem cell therapy and research

European Review ◽

10.1017/s1062798702000285 ◽

2002 ◽

Vol 10 (3) ◽

pp. 359-367 ◽

Cited By ~ 1

Author(s):

ROBIN LOVELL-BADGE

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Embryonic Stem Cells ◽

Cell Therapy ◽

Stem Cell Therapy ◽

Stem Cell Research ◽

Embryonic Stem ◽

Early Embryos ◽

Wide Range ◽

Potential Use

Stem cells are capable of regenerating tissue cells. They have an important potential use in a wide range of therapies, especially as an alternative to organ transplantation, with the advantage that they can be derived from the patient and thus avoid rejection. Embryonic stem cells are potentially capable of forming all kinds of cells. Their use is controversial however, because they are derived from early embryos and because, if they were to match the patient, they would have to be obtained using the same techniques that could, in theory, be used to produce cloned individuals. This article discusses the uses and problems of stem cell research and therapy.

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