Organs on a Chip

Readings:
Chapter 7 in Biomaterials for Clinical Applications

How can we use tissue engineering to address HIV/AIDS?

Option 1: Engineer a new immune system
Bone marrow contains hematopoietic stem cells
Individuals homozygous for CCR5 ^32 are resistant to HIV-1
stem cell transplant is curative
What are drawbacks of allogeneic (genetically different, coming from
someone else) stem cell transplant?
incredibly taxing on the patient
could potentially cause the patient to go back on
immunosuppressants, puts forth potential risk for reinfection
graft vs. host
lack of donors
expensive
benefits do not outweigh the risks
Advantages:
patient can become CCR5
do not need to find a suitable bone marrow donor
Option 2: prevent or treat with drugs and vaccines
vaccines could by prophylactic or therapeutic
small molecules & biologics could be prophylactic or therapeutic
lots of viable targets
entry
reverse transcription
chromosomal integration
maturation
small molecules do a pretty good job
Problems w/existing antiretrovirals
daily dosing
expensive
required for the rest of their lives
side effects and drug interactions
HIV-1 resistance
does not work well in developing nations
Can we make better drugs?
Drug development pathway
bench research (compound screening, lead optimization)
preclinical studies
file Investigational New Drug (IND) application w/FDAs CDER/CBER
phase 1 clinical trial: safety
 phase 2: small efficacy
phase 3: large efficacy
file new drug application (NDA) when app is pending
start manufacturing when surveillance is pending
market drugs post-market surveillance
Average time to create a new drug: 14.2 years
RO5: Rule of Five
number of hydrogen bonds
mol. weight is less that 500 Daltons
moderately liquifying

Preclinical studies
test efficacy in relevant animal model of disease
at least two animal models (one non-rodent)
disease is artificially induced (either genetically or by procedure)
identify biomarkers for clinical development
ADMET is further established in vivo
dose escalation to establish *MED and *MTD
AUC and Cmax
target organs
refine CMC/chemistry manufacturing (API manufacturing and
formulation/you know how to consistently make it)

LOA: Likelihood of Approval in Clinical Trials

How can we increase the likelihood of approval?

human 3D cell culture
human organs-on-a-chip

Traditional cell culture

immortal cell lines
cancer-derived (e.g. HeLa)
immortalized primary cells (EBV transduction)
clones, very aggresive
primary cells (e.g. HUVECS (like umbilical cords))
derived from fresh tissues
will eventually undergo replicative senescence
more genetic variability
many cells grown in two-dimensions
cells grow in monolayers on a polystyrene substrate
adherent or anchorage-dependent
some cells are anchorage-dependent
often grown in suspension
examples are PBMCs (primary) and WERI-Rb-1 (immortal)
 most human cell culture is performed at 37 degrees C
cell culture media
natural: derived from the human body
advantages: in the environment they are naturally found, happier
disadvantages:
artificial:
+/- serum: carriers, vitamins, attachment proteins, lipids, minerals,
hormones, protease inhibitors, etc.
buffer system: HEPES or bicarbonate (5-10% CO2), plus indicator
inorganic salts: sodium, potassium, and calcium
amino acids: especially L-glutamine
carbohydrates: often glucose and galactose
antibiotics (optional): protect against bacterial contamination
other cell-dependent components (e.g. estrogen)
3D cell culture
traditional cell culture does not emulate the in vivo milieu, thus results are
NOT indicative of a true physiological response
a step towards recreating tissues ex vivo
hanging drop method generates 3D spheroids
a scaffold is typically required for more advanced structures
provide support against forces (sheer stress, gravity, etc.)
offer a way to deliver nutrients
biomaterials are needed for the scaffold
synthetic polymers
poly-glycolic acid (PGA)
poly-lactic acid (PLA)
natural polymers
chitosan
collagen
What are the tradeoffs between natural and synthetic scaffolds?
synthetic are easier to modify, cheaper, more reproducible
What is missing?
movement!
spin coding- drop of liquid, spin it, even coating