CellGene Therapy by Jeff Sheehy the California Institute for Regenerative Medicine CIRM Jerome Zack UCLA HansPeter Kiem The Fred Hutchinson Cancer Research Center Jessica Handibode ID: 744105
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Slide1
Cell/gene therapy
HIV Cure Research Training Curriculum
Cell/Gene Therapy by:
Jeff Sheehy, the California Institute for Regenerative Medicine (CIRM)
Jerome Zack, UCLA
Hans-Peter
Kiem
, The Fred Hutchinson Cancer Research Center
Jessica
Handibode
,
AVAC
January, 2015
The HIV CURE training curriculum is a collaborative project aimed at making HIV cure research science accessible to the community and the HIV research field. Slide2
Session Goals/Objectives
Learn about how therapies that insert genes and use cells is on the brink of transforming medicine and curing disease.
Learn how Gene/Cell therapies fit into HIV cure efforts
Learn the targets, techniques, and cell types used in HIV Gene/Cell Therapy
Understand the risks associated with Gene/Cell therapy clinical trialsSlide3
Timothy Brown Road to a Cure for HIV
HIV+ Acute Myeloid Leukemia Patient
Identification of HLA-identical, CCR5Δ32 homozygous bone marrow donor
Chemo- and Radiotherapy Conditioning
Allogeneic stem cell transplant
6 years later: remains curedSlide4
GOOD
MORNING
AMERICA
UCLA
Researchers Announce Gene Therapy Cure for 18 ‘Bubble Baby’
Patients
Nov
18,
201418 patients with Severe Combined Immunodeficiency Disease (SCID) ranging in age from 3 months to 4 years at the time of treatment.Their blood stem cells (hematopoietic stem cells) were removed from their bone marrow and genetically modified to correct the gene defect that had left the children without a working immune system.The children were cured without any side effects.
New York TimesIn Girl’s Last Hope, Altered Immune Cells Beat LeukemiaDecember 9, 2012Juno Therapeutics, the company developing the therapy, in a study found an 89 percent remission rate among 27 adults with acute lymphoblastic leukemia no longer responding to other treatments.Doctors remove millions of the patient’s T-cells and insert new genes that enable the T-cells to kill cancer cells.The new genes program the T-cells to attack B-cells, a normal part of the immune system that turn malignant in this leukemia.The altered T-cells — called chimeric antigen receptor cells — are then dripped back into the patient’s veins, and if all goes well they multiply and start destroying the cancer.
Regenerative Medicine
/
Cell
-
Gene Therapy Maturing
Gene
modification of
patients’ own
immune cells returned to patients is saving lives.Slide5
What is Cell/Gene Therapy
A branch of
Regenerative Medicine
, an emerging field that involves
the "process of replacing, engineering or regenerating human cells, tissues or organs to restore or establish normal function”.
Gene therapy is the the
delivery of therapeutic gene into a patient's cells to treat disease.
Cell therapy
is the delivery of intact, living cells into a patient to treat disease.Combination Cell/Gene Therapy approaches that seek to insert genes into a patients’ own cells to control or kill HIV are in clinical trials now. Slide6
Ex vivo gene therapy -
Usually with blood cells
(lymphocytes
or blood stem cells) for diseases affecting the hematopoietic system
In vivo gene therapy -
Oncolytic adenoviruses for the treatment of cancer
Adeno-associated vectors for the treatment of
Duchenne
muscular dystrophy or hemophiliaNon-viral for cancer
Different Routes of Gene TherapySlide7
Sterilizing cure
complete eradication of all
replication competent forms
of HIV. The reservoir is
gone.
Timothy Brown received a sterilizing cure.
Functional Cure
Life-long control of virus
in the absence of antiretroviral therapy, but without achieving complete eradication of HIV.Virus remains in reservoirs in the body.
Cell/Gene Therapy will likely produce a functional cure, if a cure is generatedSlide8
Gene Therapy in
Blood Cells
Therapeutic HIV protection geneSlide9
Some targets for gene therapySlide10
Gene Therapy- Vectors to deliver anti-HIV genes
LV- Lentivirus vectors
RV- gammaretroviral vectors,
AAV – adeno-associated vectors
Adenovirus vectors
Vectors are replication
defective – so they cannot replicate and spread once they are inside the cells and after delivering the anti-HIV genesSlide11
Patient
Ex Vivo Gene Therapy: Putting Functional Genes Into Marrow Stem Cells or T cells Outside of the Body
Mobilization
Leukapheresis
OR
Bone Marrow Harvest
Reinfusion
Isolation of Stem Cells or T cells
Virus-Mediated Transfer of
Therapeutic Gene
GOAL: Gene modified cells engraft and correct or treat the disease
- Cancer
- Genetic disease
- Infectious diseaseSlide12
Next GenerationTechnology
Genome editing
Zinc finger
TAL Effector Nuclease
CRISPR
/
Cas9MegaTals
NH2
COOH
Zinc finger
TAL Effector Nuclease
CRISPR
/Cas9
megaTAL
Slide13
HIV target gene eg CCR5
Thanks to Barry Stoddard
Site-Specific Gene
Targeting / EngineeringSlide14
Scarless
Repair Of Genetic Defect or
Targeted Insertion Of New Genetic MaterialSlide15
Expanding gene-edited and corrected
HSCs
Development of novel
conditioning
regimens for efficient engraftment
Generation of HIV protected blood and immune system inside the patient
IIn
vivo selection
Collection HSCs
Kiem et al. Cell Stem Cell 2012 (modified)
Vector mediated gene transfer of HIV resistance genes
Nucleases for CCR5 disruption
Nucleases to eliminate integrated Virus
Patient
Patient
Hematopoietic Stem Cell Modification and
Transplantation to Cure HIV/AIDSSlide16
Current Clinical ApproachesSlide17
Timothy
Brown
--cured
of HIV
through a transplant of hematopoietic stem cells
with a natural mutation that largely prevents HIV infection. This mutation can be replicated via gene therapy.Timothy
received the stem cells from a
donor and the resulting graft
vs host disease was likely a factor in his cure. Attempts to replicate have failed in 6
patients due to the severity of their cancer.Matt Sharp took part in a clinical trial in which his own T-cells were removed from whole blood via apheresis and then gene modified and returned into his body. The Phase I trial recruited immunologic non-responders and Matt experienced a rise in his T-cell count. Sangamo, the sponsor, reported Phase II trial results in late 2014, that a “single infusion” of modified T cells “resulted in sustained reduction and control of viral load in the absence of antiretroviral drugs
in several subjects
..” and
“a decrease in the size of the HIV reservoir
.”
Cell/Gene Therapy—Why
?
One cure, human trials underwaySlide18
SANGAMO AUTOLOGOUST CELL TRIALS WITH CONDITIONING AGENT
SB-728mR-T (autologous CD4T cells genetically
Modified at the CCR5 gene) +
cyclophosphamid
NCT02225665
Phase I/II
June 2018
SB-728-T + cyclophosphamide
NCT01543152
(
closed to enrollment)
Phase I/II
Dec. 2013Slide19
Clinical trials—blood c
ancer
p
atients
Many trials recruit lymphoma or leukemia patients who need a transplant
Undergo conditioning to eliminate current immune system to create space for a new systemThe HSCs used in these trials are autologous, meaning that they are taken from the patients
not from a donor.Their HSCs are gene modified to resist HIV, and are then transplanted back into the participant in a mix of modified and unmodified cells.Slide20
Clinical t
rials-other
p
atient
populations
Other trials propose going into healthier patients—currently, either immunologic non-responders or patients who have quit taking ART (treatment fatigue) as participants.
Some of these trials include conditioning regimens which present toxicity issuesSlide21
Clinical Trial Issue
CCR5
deletion is
unlikely to be sufficient by itself in many patients.
Mutated HIV that uses the CXCR4 receptor to infect cells
is a potential complication
Gene therapy that blocks HIV in multiple ways will be needed.Slide22
Clinical Trial Issue
During
cell modification, the percentage of cells modified varies, and
a low yield
of modified cells is a barrier
.
Enough cells must be modified to achieve a therapeutic effect.
Hematopoietic cells are stimulated in a patient using drugs prior to apheresis to increase their number and percentage in the blood and enable more cells to be modified and returned. Slide23
Gene therapy clinical trial concerns
Gene therapy trials involve different gene editing/modifying techniques.
Precision is key, a serious concern is “off target” editing.
If the genes other than those targeted are modified (off target editing), the potential for serious adverse events exist, including cancer.Slide24
Treatment Interruptions
Seen as essential to allow modified cells to engraft and increase as a proportion of the cell population and to allow HIV to kill unprotected cells, and thus select for modified cells.
This process carries potential risks like
treatment regimen resistanceSlide25
Basic Science Approaches- Improving the Technology and Engineering Possible SolutionsSlide26
Patient
Expansion
of
gene-edited and
HIV protected HSCs
Collaboration Dr. Sauvageau (new UM171
molecule
Fares et al Science 2014)
Development of novel conditioning
regimens,
treosulfan
,
Astatine-211-based RIT, CAR-T cells
Generation of
genetically modified HIV protected blood
and immune system inside the patient
i
n
vivo
selection
HSC Collection
Kiem et al. Cell Stem Cell 2012 (modified)
Vector mediated
gene therapy
Nuclease-mediated
protection from HIV
Nuclease-mediated disruption of integrated HIV
Patient
Hematopoietic Stem
Cell Gene Therapy / Editing for HIVSlide27
In vivo
Selection
to increase
the Percent HIV-protected cells
O6BG/BCNU
% Gene Marking
Days After Transplantation
Gene Marking
Therapeutic ThresholdSlide28
Macrophage Activation
B-Cell function
CD8
+
T-Cell function
Cytolytic Activity
Long-term protection
Dampening of IR
Peripheral Tolerance
Maintenance of Lymphoid Tissue
Maintenance of SHIV-Specific CD4
+
T-Cells
Resistance to Direct Infection
R5- tropic
X4- tropic
Dual-tropic
Development of Gene Modified, Infection Resistant CD4
+
T-cells
Decreased Viremia
HSC M
odification Results in the Development of Infection Resistant Immune Cell Populations and an Enhanced Immune Response
Younan…Kiem Blood 2013Slide29
A genetic “handle” attached to modified cells, enabling better screening of unmodified cells
Potential purification of modified cells, reaching almost 95% purity.Slide30
Other Gene/Cell therapy approaches
The “kill” in “Kick and Kill”,
(
Lam, Baylor)
T cells are taken from the peripheral blood of patients suppressed on antiretroviral therapy.
The cells are presented with multiple HIV antigens and then expanded.
Cells are functional and have broadly specific and potent HIV infected cell killing ability and the ability to suppress HIV replication. Can be used with latency reversing agents to kill the “kicked” HIV.
HIV: Shock and Kill. Steven G
Deeks. Nature 487, 439-440 (26 July 2012)Slide31
Chimeric antigen receptor (CAR)
Antigen binding component
Expressed on outside of cell;
This can be part of an antibody, or other molecule
Usually binds HIV envelope on infected cells
HLA independent;
Signaling ComponentSends signal into the cellDirects the cell to kill HIV infected target
CD3
ζBindsViral proteinSlide32
Other approaches:
Chimeric antigen receptor T
cells (CAR T cells)
Engineering hematopoietic and T stem cells that attack and kill cells infected with HIV.
Provides a self-renewing population of both CD8+ and CD4+ HIV-targeted T-cells resistant to direct HIV infection
Also used in cancer
Jacobson, Caron A., and Jerome Ritz. "Clinical Trials Time to Put the CAR-T before the Horse."
Blood Journal
. American Society of Hematology, 3 Nov. 2011. Slide33
New avenues:In vivo gene modification
A new
class of genetic engineering tools called targeted
nucleases
make genetic engineering of stem cells much more precise and therefore safer
Deliver these reagents directly to the stem cells in the body,
Uses a viral vector that specifically targets hematopoietic cells in vivo.
HSC
T cellsSlide34
New avenues: Induced pluripotent stem cells (
iPSC
)
T
ransfer
Skin biopsy
Fibroblast reprogramming
iPSCs
generation
HIV-resistant CD4+ T cells or NK cells
HIV-resistant HSCs
Gene-modified
Skin cells are converted back into embryo-like state (
pluripotency
)
The pluripotent cells are modified to have a deletion of CCR5
Δ32 mutation
Modified cells differentiated and returnedSlide35
Conclusions
Regenerative Medicine/Cell-Gene Therapy is a rapidly maturing field offering potential for cures and therapies in several diseases and conditions
Clinical trials in HIV are underway or planned
A functional cure may result, but clinical benefit such as increased T cells for immunological non-responders would also help some patients greatly. And cell/gene therapy could provide the “kill” in “kick and kill”. It doesn’t have to lead to a cure by itself.Slide36
Conclusions
Current approaches in trial are very complex, but as the technologies develop, easier to administer and cheaper therapies will be available.
Risks, such as off-target effects and the need for treatment interruptions, are high in early trials and participants should carefully consider all risks before entering a trial.Slide37
Acknowledgements