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Cell Systems
Letter
Commendation for Exposing Key
Advantage of Organ Chip Approach
Kambez H. Benam1 and Donald E. Ingber1,2,3,*
1Wyss
Institute for Biologically Inspired Engineering at Harvard University, Boston, MA 02115, USA
John A. Paulson School of Engineering and Applied Sciences, Cambridge, MA 02139, USA
3Vascular Biology Program, Boston Childrens Hospital and Harvard Medical School, Boston, MA 02115, USA
*Correspondence: [email protected]
http://dx.doi.org/10.1016/j.cels.2016.11.009
2Harvard
Dear Reviewer #3,
We write to thank you for your help
with the review of our manuscript that
was originally entitled, Modeling normal
and COPD human lung small airway
responses to inhaled smoke in vitro,
but based on your constructive criticism,
was published last week in Cell Systems
with an entirely different title. When we
submitted this manuscript, we were
excited by our ability to engineer a robotic
cigarette smoking machine that can be
linked to a human small airway-on-achip microfluidic culture device, which
we previously showed faithfully mimics
human small airway physiology and pathophysiology. Thus, we submitted the
manuscript largely as a story focused on
development of a human lung airway
chip that smokes cigarettes. However,
because all three of the Reviewers
viewed this story from different disciplinary perspectives, and with a greater
focus on bioinformatics and human clinical studies than we had, we received
input that utterly changed how we viewed
our own story.
Specifically, your comment #2 made us
look at our own results in a different way.
You stated that, it appears that expression of only a few genes in Figure 2D
are consistent with those obtained from
human smokers. Particularly, the downregulated genes do not seem to align. A
gene of interest in validating this system
is the oxidative stress indicator HMOX1;
could the author explain the disparity in
fold change observed in normal smokers
vs. smoking chips in Figure 2D, where
many normal smokers exhibited 0 fold
change compared to the 1- to 3-fold
change observed in the chips?
This potentially publication-killing criticism made us realize that traditional human clinical study design has a major lim-
itation in that it is inherently difficult or
impossible to parse out the stimulusinduced response from background variability between individuals. For example,
in the human clinical study we used
for comparison in our manuscript, gene
expression profiles of smokers were
compared to those of average agematched non-smokers (i.e., two entirely
different patient populations), which is in
part why the range of expression levels
in normal smokers relative to non-smoking controls was so limited ( 1 to +1 fold
change). Thus, the crucial point here is
that we realized that the human airway
chips provide a major advantage over
many human clinical studies by enabling
a true matched comparison of biological
responses as the cells lining the chips
are sourced from the same individual
and then cultured in the presence or
absence of a stimulus (in this case, smoke
exposure), which normalizes for interindividual variability. In addition, in our
study, we exposed healthy donor airways
to acute smoke exposure, whereas the
clinical study compared smokers who
were chronically exposed to cigarette
smoke to healthy non-smoking donors.
Thus, the comparison to clinical data
should be expected to provide qualitative
similarity for genes related to acute
smoke exposure rather than quantitative
agreement, and this is precisely what we
observed. In addition, we discovered
that the chips reproduced the clinical
data whenever a consistent pattern was
present in over 80% of donors in the
clinical study, even with the differences
in study design between our experiments. This included expression patterns
for highly downregulated genes, as well
as for genes in the oxidation-reduction
pathway, which were upregulated in
both smoking chips and smoking individ-
uals. In context of the patient-to-patient
variability, it was also interesting to note
that individual genes, such CYP1A1,
were highly upregulated in response to
chronic smoke exposure in the clinical
samples and to acute smoke exposure
in chips, but there was much greater variability between the samples in the clinical
study.
So based on your comments, we
discovered that one of the key advantages of our organ chip approach is the
ability to perform truly matched studies
of smoke exposure, which is not possible
in vivo. In fact, the reduced variability we
obtained in our studies by performing truly
matched studies of smoke exposure
could potentially enable fewer measurements to be required to reach the same
degree of confidence for future therapeutic development.
Based on this reevaluation of our own
findings triggered by your comment, we
changed the title of our resubmitted
manuscript to Matched-Comparative
Modeling of Normal and Diseased Human
Airway Responses Using a Microengineered Breathing Lung Chip, provided
detailed point-by-point responses to all
of your concerns and those of the other
Reviewers, and convinced the Editor
to accept our manuscript for publication. This title conveys the true advance
here as we demonstrated the ability to
use human organs-on-chips to carry out
matched comparative modeling of human
responses to specific stimuli in a more
effective way than is done in many clinical
studies, and hence a better way to carry
out human pre-clinical studies in vitro.
We could not have done without you;
thank you.
Yours sincerely,
Don Ingber, MD, PhD
Kambez Benam, PhD
Cell Systems 3, November 23, 2016 2016 Published by Elsevier Inc. 411
