Cellectis Demonstrates Potential of Allogenic CAR-T Technology

at European Hematology Association (EHA), June 2015

3 communications presented by Cellectis to the European Hematology Association (EHA) 2015
demonstrate the exceptional potential of combining the CAR-T technique with the patented
Cellectis gene editing technology TALEN in the fight against 3 types of blood-hosted cancer in perclinical and mouse models. The first shows impressive results targeting the CD123 antigen in Acute
Myeloid Leukemia. The second demonstrates proof of concept in CD19+ B-cell leukemias and the
third demonstrates how TALEN elimination of the CS1 gene in CAR-T cells enhances their activity
against multiple Myeloma.
In this short document I will try to explain the methods used to develop a therapy to fight cancer
and the implications for the application to wide range of conditions. Taken together, these three
communications show just how powerful, flexible and effective this combination of technologies
can be in the fight against cancer.
PDF versions of the communications can be found at :
http://www.cellectis.com/sites/default/files/eha_2015_posters.pdf
Let's look at the specific techniques used.
The CAR component of a CAR-T cell refers to the Chimeric Antigen Receptor which is an
engineered antibody receptor that allows these cells to target antigens expressed by cancer cells.
This CAR receptor is fused to the T-cell, which when stimulated by the antigen on the cancer cell
activates cytotoxic activity to kill the targeted cancer cell. This CAR activity is common to all
CAR-T-cells which are being developed to fight against cancer. By engineering the antibody
receptor to target antigens specifically found on cancer cells, CAR-T clinical trials have shown
remission rates of 80-90% in clinical trials on refractory acute leukaemia (ALL) for which no other
options were available. Such results have given hope to cancer sufferers and promoted a rush to
develop new CAR-T therapies, but the road is still long.
A number of problems remain, one of which is cost and another is safety. On the cost side, most
treatments are based on taking blood samples from the patient himself and selecting healthy T cells
to be grown in sufficient quantities, modified to introduce the specific CAR component to target the
patient's illness before being re-injected into the patient. This personalised auto-graft approach has
the advantage that the patients immune system will not reject its own cells, but the downside is that
the process is long and costly. This restricts widespread use on economic grounds and during the
long period of processing and growing the CAR-T cells, the illness can further develop.
The use of auto-graft CAR-T cells does not however eliminate other safety issues. One of these is
that the CAR component, although targeted to antigens that are predominately found on cancer cells
may also be present (albeit in smaller quantities) on certain healthy cells. These cells will also be
targeted. The result can be side effects similar to a serious virus attack on the whole body or on
certain organs, with in some cases a massive cytokine attack causing severe inflammation, pain and
suffering. Furthermore, once the auto-graft CAR-T cells have been administered, they cannot be
deactivated, so caution must be applied to administer the best dose to attack the cancer cells without
further deteriorating the health and well-being of the patient.
The purpose of the Cellectis techniques is to give CAR-T cells the specific qualities that help to
provide additional flexibility, efficacy and safety in the fight against cancer. Some of these

techniques use the patented TALEN editing process to modify the genes in the T cells to give the
CAR-T cells qualities that ordinary T cells would not have in the absence of TALEN editing.
1) Provide an off-the-shelf treatment for each specific type of cancer.
Cellectis has pioneered the technique of using foreign donor cells from healthy patients with the
aim of creating a ready-to-use stock of CAR-T cells engineered to treat specific cancer types. This
can only be made possible if the host fails to recognise the graft T cells as foreign and accepts them
as its own cells rather than rejecting them as foreign cells. The key to this rejection is the TCR gene
which gives a signature to the graft cell's origin. Using TALEN, Cellectis edits out the TCR gene, so
that the CAR-T cell is no longer identified as foreign is not attacked by the host immune system. In
principle such cells would be immortal and protected from what is known as Graft vs Host Disease
(GvHD). The use of foreign donor cells opens the door to the development of an economically
viable range of CAR engineered T cells capable of targeting not only cancer but a whole range of
diseases including chronic viral infections.
3) Protect against self destruction/co-destruction
The CAR component built into the CAR-T cell will target an antigen found predominantly on the
targeted cancer cell. However the T cell itself carries its own antigens. When used in combination
with other treatments (chemotherapy) it may be susceptible to being attacked either by host T cells
or by combination chemotherapy agents. The TALEN gene editing technique can be used to protect
the CAR-T cells from such attack.
In the Cellectis UCART-CS1 programme, donor T cells are engineered to target the CS1 antigen
found on the surface of Multiple Myeloma cells. The allogenic (donor) T cells are first of all edited
to remove the TCR gene and so remove the risk of GvHD. However human T cells also carry the
CSI gene and so would be likely targets for self targeting by the engineered CAR-T cells. TALEN
gene editing was therefore used to remove the CSI gene from the engineered CAR-T cells to
eliminate self attack by other CAR-T cells and so potentially give the cells a longer and more
effective life.
In the UCART19 program, third party donor cells were engineered to target the CD19 antigen on Bcell Leukaemia. The TCR gene was first edited out to avoid GvHD. A common therapy for this
disease is Alemtuzumab, which targets the antigen CD52. The CD52 gene was therefore edited out
of the CAR-T cell to avoid resistance to Alemtuzumab when used in co-therapy with UCART 19.
3) Build in controlled regulation/controlled destruction
In order to modulate the activity of the CAR-T cells in case of over-reaction resulting in a violent
cytokine attack, the TALEN technique can also be used to introduce a specific suicide gene into the
CAR-T cell to render it sensitive to deactivation by a specific agent. As part of the CD123 target for
Acute Myeloid Leukemia (AML) and the UCART19 program, the T cells are designed to coexpress the RQR8 gene as a safety feature rendering them sensitive to the monoclonal antibody
rituximab.
All of these techniques were successfully demonstrated in the three communications presented by
Cellectis to the European Hematology Association (EHA) 2015. In all cases anti-tumor activity
was demonstrated in in-vivo mouse models.