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Stem Cells Off the Line

Researchers work to improve the efficiency of cell therapy manufacturing,
while developing procedures to ensure consistent quality.
BY RONNDA L. BARTEL

Over the past five years, there has been a jump in the
number of cell therapy products commercially distributed
by companies in the U.S., with eight cell
therapy products receiving approval since 2009. Once such
therapies are approved for the market, however, biopharmaceutical
companies face new challenges as they develop the
infrastructure to scale up the manufacture of larger quantities
of cell products while maintaining high standards of
quality control.
Companies advancing cell therapies at smaller scales
already face multiple challenges in executing the safe, reliable,
and consistent manufacture of a cell-based product. Crosscontamination
due to the mixing of cell samples, as well as
cell damage inflicted during freezing or other procedures, can
jeopardize the purity and efficiency of the process. Improper
labeling and record keeping could lead to a loss of patient
identity in the development of autologous treatments, which
use patient-derived cells. To reduce these risks, researchers
often use closed-system technology, in which the cells are not
exposed to the environment from the time they are put into
the system until they are delivered to the patient. This enables
large quantities of cells to be cultured at the same time; minimizes
cell manipulation to reduce the chance of contamination;
and automates the work as much as possible to curtail
human error and variability.
At Durham, North Carolina–based Argos Therapeutics,
for example, scientists have designed an automated manufacturing
process using functionally closed disposables to
meet their autologous-cell processing needs while ensuring
consistency and the ability to meet the demands of large
markets. The system involves three separate stand-alone
units that isolate and amplify RNA from a patient’s disease
sample; program dendritic cells to target disease antigens;
and perform various cellular and plasma processing steps
to generate the product. In contrast, traditional cell therapy
processes include many labor-intensive and nonintegrated
steps—including centrifugation, incubation, media addition,
cell selection, and cell washing—which require highly
skilled operators. Argos’s automated system can be rapidly
scaled up to handle large patient numbers, allowing the
company to carry out production at higher throughput levels
and with a lower overall cost of materials.
Single-use disposable bioreactors are commonly used
in a closed system to protect cell products while they are
being expanded. Using automated processes can help minimize
the risk of operator error while also ensuring sterility,
increasing productivity, and enhancing consistency of cell
batches. Robotic handling of cells in a sterile environment
has also been shown to reduce the level of particulates and
contaminants found in manually cultured cells.
Use of advanced tracking technologies can be applied to
bioreactors to further safeguard cells against a break in the
chain of identity of a patient sample, particularly when used
for autologous cells. At Aastrom Biosciences, we designed
and implemented software called Autolotrack, which automatically
assigns a lot number to each patient’s cell sample
and creates a production schedule that can be edited by the
team. This software organizes and tracks data about each
bioreactor during every phase of the production process,
and all instruments used in manufacturing automatically
feed back into the system, so that the production team can
easily see how far a cell sample has advanced in the manufacturing
process.
Preparing cells for patients
Typically, cells harvested from culture are not suitable for
direct patient administration. The cells must be processed
to remove serum and other culture reagents and to achieve
a product volume suitable for therapeutic delivery. These
steps can be time-consuming and can significantly increase
the risk of error, contamination, and excessive holding
times, as well as the potential for cell loss and decreased
viability. Autologous cell therapies pose additional challenges
for large-scale production because individual doses
must be produced for each patient.
One common step in traditional approaches to cell harvesting
is the washing away of residual culture reagents.
Bioreactors that both drain away culture medium and rinse
cells to remove residues before they are harvested eliminate
two cell-transfer steps and lead to lower rates of cell damage
and loss. Perfusion bioreactors, which continuously replace
the culture media and remove waste products, can similarly
enable cells to be harvested and purified quickly, reducing
the time dedicated to processing.

The examples listed above are just a sampling of the range
of advanced capabilities in computerization, automation, and
process integration now available to help companies drive
down costs and increase production volume and quality of
cell therapies. The application of technologies to reduce the
number of manufacturing steps or streamline overall production
should be considered at all phases of research, but especially
as a company advances to late-stage clinical trials and
prepares for commercialization. Success requires continually
identifying and assessing viable advances in technology
that can be applied to any phase of cell therapy production
and incorporating such advances at the earliest opportunity to
improve performance and reduce risk. 􀁊
Ronnda L. Bartel is the chief scientific officer at Aastrom
Biosciences, a Michigan-based biotechnology company focused
on patient-specific cell therapy applications.


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Fighting Cancer with Nanomedicine

Nanotechnology-based therapeutics will revolutionize cancer treatment.
BY DEAN HO

Short drug circulation times and difficulty localizing therapy
to tumor sites are but two of the challenges associated
with existing cancer treatments. More troubling are
the issues of drug toxicity and tumor resistance. Toxicity can
cause major complications, such as low white-blood-cell counts
or heart fail ure, that necessitate cessation of treatment. The
tissue damage inflicted by some therapies can even be fatal.
And evolution of drug resistance by tumors accounts for the
vast majority of cases in which treatment fails. Given these and
other issues associated with treatment safety and efficacy, scientists
are applying tremendous effort toward the utilization of
nanomedicine in the fight against cancer.
Nanotechnology-based therapeutics have exhibited clear
benefits when compared with unmodified drugs, including
improved half-lives, retention, and targeting efficiency, and
fewer patient side effects. Researchers have already made progress
with chemotherapeutic nanomedicines in the clinic. Several
compounds that are in various stages of trials or already
approved by the U.S. Food and Drug Administration (FDA).
For example, Calando Pharmaceuticals has demonstrated the
first evidence of nanoparticle-delivered clinical RNA interference
(RNAi) (Nature, 464:1067-70, 2010). BIND Biosciences
has shown that nanoparticles combining a chemotherapeutic
drug with prostate-specific membrane antigen (PSMA) can
reduce lung and tonsillar lesions with greater efficacy compared
with the drug alone, and at substantially lower doses (Sci Transl
Med, doi:10.1126/scitranslmed.3003651, 2012). Furthermore
Celgene’s Abraxane, an albumin-functionalized paclitaxel formulation,
was initially approved by the FDA for sale as a breast
cancer therapy, but also recently received approval for the treatment
of lung and pancreatic cancers.
On the preclinical front, several nanomaterial formulations have
shown promise. Single-agent nanoparticle delivery, both actively and
passively targeted, has been demonstrated with a host of platforms
using silica, polymer, metal, and carbon-based materials.
Delivering a double whammy
Researchers recently reported multidrug delivery using nanoparticles
to mediate resistance in relapsing cancers and to improve
triple-negative breast cancer treatment efficacy. Other recent
approaches have included layer-by-layer siRNA and doxorubicin
delivery for breast cancer therapy, simultaneous loading of
small interfering RNA (siRNA) and tumor-penetrating peptides
against ovarian cancer, as well as sequential administration of multiple
types of nanoparticles for pancreatic cancer treatment (Adv
Funct Mater, doi:10.1002/adfm.201303222, 2014). These exciting
approaches have served as a foundation for the next phase of cancer
nanomedicine in the clinic—the rational design of nanomaterial-
drug combinations.
Until more nanoparticles are validated in the clinic, however,
the impact that nanomedicine may have on cancer treatment
has yet to be fully realized. In order for chemotherapies
modified using nanotechnology to profoundly change hematological
and oncological practice, the application of engineered
nanomedicines must be paired with emerging strategies to
rationally design nanotherapeutic combinations. This is critical
because combinatorial therapy is an efficient way to simultaneously
address the barriers to treatment success, and it is
widely used in treating cancer and infectious diseases.
Current clinical methodologies for combinatorial drug design
include additive treatments that combine two or more drugs
at their highest tolerable but still efficacious dose, although the
synergistic effects among drugs cannot be taken into account
using this additive approach. As the field gradually embraces the
use of nanoparticles to deliver multiple compounds with different
targets, a move away from additive dosing is necessary. This
raises several important questions. For example, silencing genes
to combat resistance, mediating apoptosis, and allowing vascular
access are each pathways worth targeting, but what if multiple
pathways are targeted at the same time to comprehensively
attack the tumor? How will dosing be determined? How will the
dosages of each drug be adjusted if efficacy is improved but toxicity
is worsened? More importantly, how will “optimization” be
defined, especially if the desired outcome is to simultaneously
stop tumor growth, eliminate resistance, maintain white blood
cell counts, and achieve a host of other objectives?
An attempt to optimize any one of these conditions will inevitably
affect the others. Furthermore, these conditions vary from
patient to patient, so phenotypic personalized medicine will be
required. In addition, these issues create a parameter space that
is too large to be individually tested and can result in an arbitrary
dosing scenario. For example, a combination of six candidate
therapeutics with 10 possible concentrations represents a minimum
of 1 million possible combinations. Identifying a solution
that rapidly converges on a defined set of phenotypic outcomes is
a challenge that faces both unmodified drug administration and
drug delivery by nanoparticles.
To move beyond short-term cancer management—or single
outcomes, like delaying tumor growth using a nanoparticle drug
formulation—and to enable long-term or potentially permanent
disease management, the field of nanomedicine will inevitably
need to be paired with advanced strategies to rapidly determine
dosing conditions that can simultaneously optimize for
efficacy and safety. One promising route is the field of feedback
system control (FSC), which relies on phenotypic responses
instead of trying to interrogate cellular pathways, their individual
protein components, or a spectrum of genotypic responses.
One example is the use of a search algorithm in a feedback loop
that can guide the formulation of rational drug combinations,
both unmodified and nanotherapeutic. (See PNAS, 105:5105-
10, 2008; BMC Systems Biology, 5:88, 2011.) Remarkably, this
approach can be used for in vitro studies with cell lines and primary
cells, and for preclinical and even clinical validation. And
because FSC utilizes outcomes to iteratively suggest new possible
combinations before rapid convergence—in tens of trials
versus a million or more—toward an optimal combinatorial
dose, pharmacokinetics and pharmacodynamics are inherently
accounted for with this approach. Furthermore, because combinations
will vary from patient to patient, FSC will help personalized
nanomedicine dosing on a case-by-case basis.
In sum, cancer nanomedicine possesses the versatility
required to uniquely overcome some of the most challenging
impediments to treatment success. Rationally designing
nanotherapeutic combinations using rapid convergence solutions
such as FSC represents a promising pathway from cancer
management towards cancer elimination. 􀁊
Dean Ho is a professor of oral biology and medicine at
the University of California, Los Angeles (UCLA) School
of Dentistry, where he codirects the Weintraub Center for
Reconstructive Biotechnology. He is also a UCLA professor of
bioengineering and a member of the Jonsson Comprehensive
Cancer Center and California NanoSystems Institute.


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