Clinical scale zinc finger nuclease mediated gene editing of PD-1 in tumor infiltrating lymphocytes for the treatment of metastatic melanoma.

Several Sangamo BioSciences authors contributed to this important research.
Clinical scale zinc finger nuclease mediated gene editing of PD-1 in tumor infiltrating lymphocytes for the treatment of metastatic melanoma.
Beane JD¹, Lee G², Zheng Z³, Mendel M², Abate-Daga D⁴, Bharathan M³, Black M³, Gandhi N², Yu Z³, Chandran S³, Giedlin M², Ando D², Miller J², Paschon D², Guschin D², Rebar EJ², Reik A², Holmes MC², Gregory PD², P Restifo N³, Rosenberg SA³, Morgan RA⁵, Feldman SA³.

Abstract

Programmed cell death-1 (PD-1) is expressed on activated T cells and represents an attractive target for gene-editing of tumor targeted T cells prior to adoptive cell transfer (ACT). We used zinc finger nucleases (ZFNs) directed against the gene encoding human PD-1 (PDCD-1) to gene-edit melanoma TIL. We show that our clinical scale TIL production process yielded efficient modification of the PD-1 gene locus, with an average modification frequency of 74.8% (n=3, range 69.9 - 84.1%) of the alleles in a bulk TIL population, which resulted in a 76% reduction in PD-1 surface-expression. Forty to 48% of PD-1 gene-edited cells had biallelic PD-1 modification. Importantly, the PD-1 gene-edited TIL product showed improved in vitro effector function and a significantly increased polyfunctional cytokine profile (TNFα, GM-CSF and IFNγ) compared to unmodified TIL in two of the three donors tested. In addition, all donor cells displayed an effector memory phenotype and expanded approximately 500 - 2000 fold in vitro. Thus, further study to determine the efficiency and safety of adoptive cell transfer using PD-1 gene-edited TIL for the treatment of metastatic melanoma is warranted.Molecular Therapy (2015); doi:10.1038/mt.2015.71.

Role of the macrophage in HIV-associated neurocognitive disorders and other comorbidities in patients on effective antiretroviral treatment

Abstract

Combination antiretroviral therapy (ART) has altered the outcomes of HIV infection in treated populations by greatly reducing the incidence of opportunistic infections, cancer, and HIV-associated dementia. Despite these benefits, treated patients remain at high risk of chronic diseases affecting the peripheral organs and brain. Generally, these morbidities are attributed to persistence of latent HIV in resting T cells, chronic inflammation, and metabolic effects of ART. This review makes the case that monocytes/macrophages warrant attention as persistent reservoirs of HIV under ART, source of systemic and brain inflammation, and important targets for HIV eradication to control chronic HIV diseases.

 

 

Journal of NeuroVirology      

Date: 02 May 2015

GSK files for Gene Therapy approval in Europe

From Reuters:
http://www.reuters.com/article/2015/05/05/us-gsk-genetherapy-idUSKBN0NQ0TR20150505

GlaxoSmithKline said on Tuesday it had submitted a gene therapy for approval in Europe, becoming the first big drugmaker to seek marketing authorization for the technology to fix faulty genes.
Reuters reported last week that the move was imminent. It marks the latest sign of a renaissance in gene therapy after some disastrous clinical trial results in the late 1990s and early 2000s.
GSK's product, developed with Italian scientists, is designed to treat a tiny number of children with ADA Severe Combined Immune Deficiency (ADA-SCID) for whom no suitable bone marrow donor can be found.

U PENN Recieves $7.5 million grant from NIH for UPENN/Sangamo HIV research

Penn Medicine Researchers Receive $7.5 Million to Expand HIV Gene Therapy Work

Penn-led team will engineer T cells to be resistant to HIV-1 infection

PHILADELPHIA – Researchers from the Perelman School of Medicine and the Penn Center for AIDS Research (CFAR) have been awarded $7.5 million over five years from the National Institutes of Health to initiate a multi-project HIV study investigating a new gene therapy approach to render immune cells of HIV positive patients resistant to the virus.
The project, entitled “Engineering T cells to Provide Durable Control of HIV-1 Replication,” includes principal investigator James L. Riley, PhD, associate professor of Microbiology, Pablo Tebas, MD, director of the AIDS Clinical Trials Unit at the Penn CFAR, James Hoxie, MD, professor of Medicine at Penn and director of the Penn CFAR, Michael C. Holmes, PhD, VP Research at Sangamo BioSciences Inc.,  E. John Wherry, PhD, professor of Microbiology and director of the Institute for Immunology, and  Frederick D. Bushman, PhD, professor of Microbiology.
The Penn-led team, in collaboration with Sangamo, will investigate the ability of a synthetic molecule consisting of a viral entry inhibitor called C34 fused to CXCR4, an HIV co-receptor used by the virus to enter and infect T cells.  Building upon the success of past studies that utilized a zinc finger nuclease (ZFN) technology to disrupt the other major viral entry factor, CCR5, the new Penn project—in both preclinical and clinical studies—aims to safely build an army of modified, HIV-1 resistant T cells in HIV infected patients using a lentiviral technology to express the C34-CXCR4 molecule. This approach, researchers believe, will make more CD4 T cells resistant to the virus and thus may re-invigorate the immune response to control HIV-1 replication in the absence of antiretroviral drug therapy (ADT).
The grant is funded under NIH’s U19 Research Program, which funds collaborative projects involving multiple institutions, including an industry collaborator. The purpose of this funding opportunity, supported by the National Institute of Allergy and Infectious Disease, National Heart, Lung, and Blood Institute, and the National Institute of Mental Health, is to encourage investigators to work together on innovative approaches to eliminate HIV and leverage the expertise and resources of the participating institutes.
For the preclinical work on the Penn project, researchers will learn more about how the mechanism by C34-CXCR4 provides such robust protection of CD4 T cells and apply chimeric antigen receptor (CAR) technology to re-direct the HIV-1 immune response, among other laboratory efforts. As part of the phase I study, researchers will manufacture the engineered T cells (taken from a patients’ own immune system) and then infuse them back into HIV infected patients taken off ADT to determine their resistance and survival abilities and anti-viral effects.
The work is an important extension of the HIV gene therapy work conducted by researchers at the Penn CFAR and the Perelman School of Medicine over the last decade.
Most recently, reporting in the New England Journal of Medicine in 2014, Penn researchers successfully genetically engineered the immune cells of 12 HIV positive patients to resist infection, and decreased the viral loads of some patients taken off ADT entirely—including one patient whose levels became undetectable. The group used the ZFN technology developed by Sangamo to modify the T cells in the patients—a “molecular scissors,” of sorts, to mimic the CCR5-delta-32 mutation. That rare mutation is of interest because it provides a natural resistance to the virus, but in only 1 percent of the general population. By inducing the mutations, the scientists reduced the expression of CCR5 surface proteins. Without those, HIV cannot enter, rendering the patients’ cells resistant to infection. In April 2015, the researchers received the National Clinical Research Achievement Award for the paper.
 “In this next trial, we hypothesize that we will observe a significant loss in viral load for a longer period of time because we are able to protect CD4 T cells from different classes of HIV with the C34-CXCR4 approach,” Riley said. “This is an important component of the HIV gene therapy work here at Penn, as it will tell us how these approaches stack up against each other. We are ultimately trying to find effective, lasting ways to eliminate the need for lifelong ADT for HIV infected patients, and we believe these projects will help find the answers to some of the hard questions surrounding that goal.”
The NIH grant number for this project is U19 AI117950.

Dale Ando earns prestigious FDA appointment

Dale Ando M.D., Vice President Therapeutic Development and Chief Medical Officer of Sangamo BioSciences (SGMO) has been appointed to the Cellular, Tissue and Gene Therapies Advisory Committee of the FDA. His term runs from 4/1/2015 to 3/31/2019. His area of expertise is Industry Representative.

John Zaia, "The Goal is a Functional Cure"

In an interview in the New Scientist Magazine, principal investigator for the soon to begin Sangamo BioSciences (SGMO) stem cell HIV trial John Zaia states " The Goal is a Functional Cure".
John Zaia, M.D. is the  Institutional Official for Research, City of Hope, Duarte CA.
It is great to hear him speak with such confidence.
The article also touches on the upcoming sickle cell  trial:
"The trial blazes a path for using the approach to treat other diseases. For example, another trial set to start soon will focus on sickle cell disease, in which the oxygen-carrying haemoglobin molecules in red blood cells are abnormal. The technique would switch on a protein that can be used instead of the haemoglobin."
http://www.newscientist.com/article/mg22630194.200-human-gene-editing-has-arrived--heres-why-it-matters.html?full=true#.VUS-YZ0o7vZ

Genomeweb review of CAR T-cell Therapies: Sangamo Mention

The
Genomeweb website has an excellent review of the CAR-T landscape. It also delves into the benefits of various gene editing techniques. The article can be found here:
https://www.genomeweb.com/gene-silencinggene-editing/gene-editing-drives-development-cancer-t-cell-therapies
Here are some relevant quotes:
With numerous receptors to target, CAR T cells could be engineered every which way to target many kinds of cancer — there's even evidence they could be used to treat solid tumors. But so far they only work as an autologous treatment, where the T cells are isolated from the patient, engineered ad hoc, grown to great numbers, and reintroduced to that same patient. Otherwise, if the T cells came directly from another patient, they would likely turn against the body, inducing graft versus host disease (GVHD).
"From an operational standpoint, it's almost a nightmare," Friesen said of autologous CAR T cell treatment. "It's very difficult and commercially that's not a very viable proposition."

Both Janssen and Cellectis, a Paris-based firm that recently opened a CAR T-cell research center in New York, are pursuing development of an off-the-shelf, or allogeneic, CAR T-cell product that would not cause GHVD. To do this, they'll need to engineer cells so that they don't attack the cells in the body and vice versa, and to make it happen they've both made bold decisions on genome-editing technologies.  Janssen recently partnered with Transposagen to use CRISPR/Cas9, while Cellectis is going a different route with transcription activator-like effector nucleases (TALENs).

Other companies that have announced CAR T cell projects include Kite, Bluebird, and Sangamo Biosciences. Sangamo is trying to engineer T cells to eliminate HIV, using its exclusive zinc finger nuclease gene-editing technology.

But Janssen — and anybody else who is using CRISPR to engineer T cells — may run into problems, André Choulika, CEO of Cellectis, told GenomeWeb.
"It's a new technology for underfunded academic labs, but it's not a technology for industrial therapeutic labs," he said. "The technology is not ready yet."
Cellectis plans to use TALENs in its CAR T cell development, a technology for which it has acquired an exclusive license from the University of Minnesota. Dan Voyntas, a professor there and inventor of TALENs, is the head of Cellectis' plant sciences division.
But Choulika said the company has done its due diligence and prefers TALENs for a variety of reasons. "We've tried them all — Meganucleases, ZFNs, CRISPR. We benchmarked them, comparing all the different technologies," he said, adding that TALENs come out on top in a number of important metrics for engineering CAR T cells.
Choulika said Cellectis can routinely get TCR alpha knockout efficiency of 80 to 95 percent with TALENs. Cellectis measured CRISPR/Cas9 knockout efficiency at around 25 percent, thus any cost savings would be partially lost due to lower effectiveness.
Another problem at this stage in CRISPR/Cas9 technology development is the off-target effects. "You can get your knockouts, but the T cell will lose one thing, which is t-cell expansion. Most of the cells won't be able to amplify," Choulika said.