Investigating Optimal Autologous Cellular Platforms for Prenatal or Perinatal Factor VIII Delivery to Treat Hemophilia A

Stem, Christopher; Rodman, Christopher; Ramamurthy, Ritu M; George, Sunil; Meares, Diane; Farland, Andrew M.; Atala, Anthony; Doering, Christopher B.; Spencer, H. Trent; Porada, Christopher D.; Almeida-Porada, Graça

doi:10.3389/fcell.2021.678117

Cited by 4 publications

(7 citation statements)

References 103 publications

(128 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With regard to tissue-resident EPCs, pre-cultured human CD31 + c-Kit + VEGFR + cells isolated form umbilical cord tissue (UCT) EPCs have been shown to express similar levels of F8 gene mRNA as cultured hLSECs. Moreover, retroviral-FVIII-transduced UCT-EPCs have proved to be highly amenable to expressing vector-encoded FVIII, producing levels of mRNA, protein, and procoagulant activity that far exceeded those of transduced LSECs, which are thought to be the body’s primary site of FVIII synthesis [ 203 ].…”

Section: Clotting Factor Biosynthesismentioning

confidence: 99%

The Vascular Endothelium and Coagulation: Homeostasis, Disease, and Treatment, with a Focus on the Von Willebrand Factor and Factors VIII and V

Pablo-Moreno

Serrano

Revuelta

et al. 2022

IJMS

View full text Add to dashboard Cite

The vascular endothelium has several important functions, including hemostasis. The homeostasis of hemostasis is based on a fine balance between procoagulant and anticoagulant proteins and between fibrinolytic and antifibrinolytic ones. Coagulopathies are characterized by a mutation-induced alteration of the function of certain coagulation factors or by a disturbed balance between the mechanisms responsible for regulating coagulation. Homeostatic therapies consist in replacement and nonreplacement treatments or in the administration of antifibrinolytic agents. Rebalancing products reestablish hemostasis by inhibiting natural anticoagulant pathways. These agents include monoclonal antibodies, such as concizumab and marstacimab, which target the tissue factor pathway inhibitor; interfering RNA therapies, such as fitusiran, which targets antithrombin III; and protease inhibitors, such as serpinPC, which targets active protein C. In cases of thrombophilia (deficiency of protein C, protein S, or factor V Leiden), treatment may consist in direct oral anticoagulants, replacement therapy (plasma or recombinant ADAMTS13) in cases of a congenital deficiency of ADAMTS13, or immunomodulators (prednisone) if the thrombophilia is autoimmune. Monoclonal-antibody-based anti-vWF immunotherapy (caplacizumab) is used in the context of severe thrombophilia, regardless of the cause of the disorder. In cases of disseminated intravascular coagulation, the treatment of choice consists in administration of antifibrinolytics, all-trans-retinoic acid, and recombinant soluble human thrombomodulin.

show abstract

Section: Clotting Factor Biosynthesismentioning

confidence: 99%

The Vascular Endothelium and Coagulation: Homeostasis, Disease, and Treatment, with a Focus on the Von Willebrand Factor and Factors VIII and V

Pablo-Moreno

Serrano

Revuelta

et al. 2022

IJMS

View full text Add to dashboard Cite

show abstract

“…This is especially true for the hLSECs, which exhibit very slow division kinetics and cannot be propagated for more than a couple of passages in vitro prior to senescing. The PLCs employed in the present study exhibit a phenotype and biological properties that closely resemble that of MSC from other tissues ( 3 , 22 , 68 ). It is noteworthy that prior studies have reported the downregulation of components of the DNA damage response (DDR) and HRR pathways in MSC with time in culture ( 94 – 96 ).…”

Section: Discussionmentioning

confidence: 87%

“…While current clinical trials are employing direct injection of AAV-based vectors (7)(8)(9)(10)(11)(12)(13), the use of gene-modified cells as vehicles to accomplish gene "addition" has many advantages from a manufacturing standpoint, as it allows multiple safeguards to be added to the production process (57). In an effort to make cell-based gene therapy a clinical reality for HA, we and others have performed studies over the past decades to identify the ideal cell type for delivering a fVIII transgene and the optimal vector to introduce the fVIII transgene into the desired cell population (3,22,31,34,(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48). Previously, we demonstrated the advantages of using lentivector-transduced PLCs as the cellular vehicle for delivering a fVIII transgene (22,68).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Comparison of different gene addition strategies to modify placental derived-mesenchymal stromal cells to produce FVIII

et al. 2022

Self Cite

View full text Add to dashboard Cite

IntroductionPlacenta-derived mesenchymal cells (PLCs) endogenously produce FVIII, which makes them ideally suited for cell-based fVIII gene delivery. We have previously reported that human PLCs can be efficiently modified with a lentiviral vector encoding a bioengineered, expression/secretion-optimized fVIII transgene (ET3) and durably produce clinically relevant levels of functionally active FVIII. The objective of the present study was to investigate whether CRISPR/Cas9 can be used to achieve location-specific insertion of a fVIII transgene into a genomic safe harbor, thereby eliminating the potential risks arising from the semi-random genomic integration inherent to lentiviral vectors. We hypothesized this approach would improve the safety of the PLC-based gene delivery platform and might also enhance the therapeutic effect by eliminating chromatin-related transgene silencing.MethodsWe used CRISPR/Cas9 to attempt to insert the bioengineered fVIII transgene “lcoET3” into the AAVS1 site of PLCs (CRISPR-lcoET3) and determined their subsequent levels of FVIII production, comparing results with this approach to those achieved using lentivector transduction (LV-lcoET3) and plasmid transfection (Plasmid-lcoET3). In addition, since liver-derived sinusoidal endothelial cells (LSECs) are the native site of FVIII production in the body, we also performed parallel studies in human (h)LSECs).ResultsPLCs and hLSECs can both be transduced (LV-lcoET3) with very high efficiency and produce high levels of biologically active FVIII. Surprisingly, both cell types were largely refractory to CRISPR/Cas9-mediated knockin of the lcoET3 fVIII transgene in the AAVS1 genome locus. However, successful insertion of an RFP reporter into this locus using an identical procedure suggests the failure to achieve knockin of the lcoET3 expression cassette at this site is likely a function of its large size. Importantly, using plasmids, alone or to introduce the CRISPR/Cas9 “machinery”, resulted in dramatic upregulation of TLR 3, TLR 7, and BiP in PLCs, compromising their unique immune-inertness.DiscussionAlthough we did not achieve our primary objective, our results validate the utility of both PLCs and hLSECs as cell-based delivery vehicles for a fVIII transgene, and they highlight the hurdles that remain to be overcome before primary human cells can be gene-edited with sufficient efficiency for use in cell-based gene therapy to treat HA.

show abstract

“…18 93 94 95 Moreover, retroviral FVIII-transduced human umbilical cord-derived ECFCs are superior to transduced LSECs (liver sinusoidal endothelial cells) in terms of mRNA production, protein levels, and procoagulant activity. 96…”

Section: Ecfcs For Potential Therapeutic Applicationsmentioning

confidence: 99%

Potentials of Endothelial Colony-Forming Cells: Applications in Hemostasis and Thrombosis Disorders, from Unveiling Disease Pathophysiology to Cell Therapy

Schwarz,

Yadegari

2023

Hamostaseologie

View full text Add to dashboard Cite

Endothelial colony-forming cells (ECFCs) are endothelial progenitor cells circulating in a limited number in peripheral blood. They can give rise to mature endothelial cells (ECs) and, with intrinsically high proliferative potency, contribute to forming new blood vessels and restoring the damaged endothelium in vivo. ECFCs can be isolated from peripheral blood or umbilical cord and cultured to generate large amounts of autologous ECs in vitro. Upon differentiation in culture, ECFCs are excellent surrogates for mature ECs showing the same phenotypic, genotypic, and functional features. In the last two decades, the ECFCs from various vascular disease patients have been widely used to study the diseases' pathophysiology ex vivo and develop cell-based therapeutic approaches, including vascular regenerative therapy, tissue engineering, and gene therapy. In the current review, we will provide an updated overview of past studies, which have used ECFCs to elucidate the molecular mechanisms underlying the pathogenesis of hemostatic disorders in basic research. Additionally, we summarize preceding studies demonstrating the utility of ECFCs as cellular tools for diagnostic or therapeutic clinical applications in thrombosis and hemostasis.

show abstract

Investigating Optimal Autologous Cellular Platforms for Prenatal or Perinatal Factor VIII Delivery to Treat Hemophilia A

Cited by 4 publications

References 103 publications

The Vascular Endothelium and Coagulation: Homeostasis, Disease, and Treatment, with a Focus on the Von Willebrand Factor and Factors VIII and V

The Vascular Endothelium and Coagulation: Homeostasis, Disease, and Treatment, with a Focus on the Von Willebrand Factor and Factors VIII and V

Comparison of different gene addition strategies to modify placental derived-mesenchymal stromal cells to produce FVIII

Potentials of Endothelial Colony-Forming Cells: Applications in Hemostasis and Thrombosis Disorders, from Unveiling Disease Pathophysiology to Cell Therapy

Contact Info

Product

Resources

About