Compounds for improved viral transduction
Inventors
Heffner, Garrett Collins • Bassan, Abraham Isaac
Assignees
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Publication Number
US-10907177-B2
Publication Date
2021-02-02
Expiration Date
Abstract
The present invention provides methods and compositions for improving efficacy of viral transduction of cells. More particularly, the present invention provides methods and materials useful for safely and reliably improving the efficiency of methods and materials useful for safely and reliably improving the efficiency of methods for transducing cells, such as human hematopoietic stem cells (HSC), with viruses and/or viral vectors. The compositions and methods are useful for therapeutic indications amenable to treatment with hematopoietic stem cell gene therapies.
Core Innovation
The disclosed invention relates to gene therapy approaches for increasing the lentiviral vector transduction efficiency of CD34+ hematopoietic stem or progenitor cells. The improvement is achieved by ex vivo contacting the CD34+ hematopoietic stem or progenitor cells with a culture medium comprising a lentiviral vector and prostaglandin E2 (PGE2), 16,16-dimethyl PGE2, or an analogue thereof, where the lentiviral vector transduction efficiency is increased compared to contacting with a culture medium comprising the lentiviral vector in the absence of the prostaglandin.
The document further describes that prostaglandin EP receptor signaling is increased by using PGE2, 16,16-dimethyl PGE2, or EP ligands, including EP receptor agonists and analogues or derivatives. The disclosed framework optionally includes combining the prostaglandin condition with histone deacetylase (HDAC) inhibitors in the culture medium to improve transduction efficiency.
In addition, the document provides disclosed classes and architectures of lentiviral vectors for use with the prostaglandin-containing culture medium. The lentiviral vectors include retro/lentiviruses, including HIV-1-derived vectors, replication-defective/SIN designs, and VSV-G pseudotyping, and the disclosure includes exemplary regulatory and packaging elements such as HIV-1 LTR components, Ψ (Psi) packaging signal, cPPT/FLAP, RRE, and β-globin promoter/LCR, along with a 3′ SIN LTR including insulator elements or a rabbit β-globin polyA (rβgpA) sequence.
The disclosure additionally includes experimental support focused on measuring transduction efficiency outcomes such as GFP+ percentages and vector copy number, together with downstream validation described for CD34+ hematopoietic stem/progenitor cell applications. The document further describes additional analyses including integration site analysis and functional evaluation in downstream models described in the disclosure.
Claims Coverage
The partial content includes three independent claims directed to increasing lentiviral transduction efficiency of CD34+ hematopoietic stem or progenitor cells by ex vivo contacting with a lentiviral vector plus either PGE2/an analogue, PGE2 specifically, or 16,16-dimethyl PGE2. Across these independent claims, the core coverage centers on prostaglandin-containing culture media yielding increased lentiviral transduction efficiency compared to the same culture medium lacking the specified prostaglandin.
Prostaglandin E2 increases lentiviral transduction efficiency
Contacting CD34+ hematopoietic stem or progenitor cells with a culture medium comprising a lentiviral vector and prostaglandin E2 (PGE2), 16,16-dimethyl PGE2, or an analogue thereof ex vivo, wherein the lentiviral vector transduction efficiency is increased compared to CD34+ hematopoietic stem or progenitor cells contacted with the same culture medium comprising the lentiviral vector in the absence of PGE2, 16,16-dimethyl PGE2, or an analogue thereof.
Prostaglandin E2 increases lentiviral transduction efficiency (PGE2-containing medium)
Contacting CD34+ hematopoietic stem or progenitor cells with a culture medium comprising a lentiviral vector and PGE2 ex vivo, wherein the lentiviral transduction efficiency is increased compared to CD34+ hematopoietic stem or progenitor cells contacted with the same culture medium comprising the lentiviral vector in the absence of PGE2.
16,16-dimethyl PGE2 increases lentiviral transduction efficiency
Contacting CD34+ hematopoietic stem or progenitor cells with a culture medium comprising a lentiviral vector and 16,16-dimethyl PGE2 ex vivo, wherein the lentiviral transduction efficiency is increased compared to CD34+ hematopoietic stem or progenitor cells contacted with the same culture medium comprising the lentiviral vector in the absence of 16,16-dimethyl PGE2.
The independent claims collectively cover ex vivo methods for enhancing lentiviral transduction of CD34+ hematopoietic stem/progenitor cells by adding PGE2, 16,16-dimethyl PGE2, or an analogue thereof to the culture medium, with the increase defined relative to conditions lacking the specified prostaglandin.
Stated Advantages
Increased lentiviral vector transduction efficiency of CD34+ hematopoietic stem or progenitor cells when contacted with a prostaglandin-containing culture medium compared to the absence of the prostaglandin.
Documented Applications
Gene therapy for hemoglobinopathies, including β-thalassemia and sickle cell disease, using lentiviral vector transduction of human hematopoietic stem/progenitor cells with prostaglandin EP receptor signaling.
Gene therapy for adrenoleukodystrophy/adrenomyeloneuropathy using lentiviral vector transduction of CD34+ hematopoietic stem or progenitor cells with prostaglandin EP receptor signaling.
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