Journal articles: (1) “A Systems Biological View of Life-and-Death Decision with Respect to

Endoplasmic Reticulum Stress-The Role of PERK Pathway” (2) “Protein misfolding in the endoplasmic
reticulum as a conduit to human disease” (3) “Potential for therapeutic manipulation of the UPR in
disease”

Unfolding Protein Response (UPR) activation in the Endoplasmic Reticulum as a deciding factor
to alleviate or initiate human diseases.

Thesis Statement: Marton et al. (2017) gave deep analysis on the mechanism of UPR as how it is
induced. Furthermore, Wang & Kaufman (2016) discussed that overactivation of UPR due to ER stress by
environmental influence or increased protein synthesis affects the onset of related diseases; however,
Park & Ozcan (2013) suggested that understanding how UPR works has a potential to be therapeutically
beneficial.

Introduction:

The endoplasmic reticulum (ER) is a vital organelle since it functions as the protein and lipid
factory inside a eukaryotic cell. But synthesis of these two biomolecules occur in separate type of
organelles; rough ER for protein synthesis, and smooth ER for lipid production. Protein that is being
synthesized in the rough ER includes secretory and membrane bound proteins. Rough ER, aside from
protein synthesis, it also plays a role in protein folding and quality control. In protein folding, RER
regulates the protein folding homeostasis. Protein folding homeostasis acts as a determining factor
whether the ER is under stress. According to Wang & Kaufman (2016), without protein folding
homeostasis this may suggest that ER stress would be a major contributor to the development of various
pathological diseases. Other factors such as, increased protein synthesis, genetic mutations that causes
protein misfoldings, alteration in calcium homeostasis, and nutrient starvation such as glucose
deprivation. To recuperate from ER stress, the cell initiates a defense mechanism called unfolded protein
response (UPR).

Unfolded response protein (UPR) function as a regulator that maintains ER homeostasis when
the cell is undergoing ER stress. UPR is also activated due to environmental insults may lead to protein
misfolding, an ER stress. Accumulation of ER stress activates the three branches of UPR which will drive
back the cell to homeostatic state by self-eating dependent autophagy but over-production of UPR can
trigger apoptosis, which leads to cell death. This process, UPR, is initiated by three major transducers:
protein ER kinase (PERK), inositol requiring enzyme-1 (IRE1), and activating transcription factor-6
(ATF6). These three branches of UPR can be activated through different processes to mediate/alleviate
ER stress in the cell. According to Marton et al. (2016) PERK, under ER stress, reduces protein synthesis
which decreases the speed of entry of new polypeptides into the ER. This facilitates the homeostasis of
protein folding, thus enabling the unfolded protein in the lumen to be in their folded state. IRE1, when
activated by homodimerization during ER stress, induces its RNase activity that enables the initiation of
unconventional splicing of a transcription factor, X-box binding protein 1 (XBP1) which promotes cell
survival by regulating the transcription of various genes that is involved in protein folding, protein quality
control, phospholipid synthesis, and ER-associated degradation (ERAD). IRE1 is also responsible of
activation of apoptotic cell death due to excessive amount of XBP1. And the last branch of UPR, ATF6
when activated during ER stress, AFT6 is transported to Golgi bodies and is introduced to proteolysis and
is cleaved. The cleaved ATF6 acts as a transcription factor that will activate the UPR target genes. These
three branches of UPR dictates whether UPR is activated.

Now how can UPR determine whether it can help in alleviating of diseases or help in worsening
the patient’s disease? ER stress activates UPR and UPR initiates cell death by apoptosis or UPR
promotes protein folding homeostasis which lessens protein misfoldings that would be transported
outside RER and in to Golgi bodies. Studies conducted by Park and Ozcan (2013) indicated
advancement in the regulation of UPR and its branches in order to develop strategies in treating
diseases. UPR balances the protein folding homeostasis thus reducing the effects of ER stress in
diseases because intensity of ER stress in the cell indicates the intensity and complexity of the disease.
Reduction the effects of ER stress may vary based on the specific disease and may be very specific
depending on the disease.

Discussion:

The endoplasmic reticulum (ER) is a specialized organelle that has crucial roles in cell
homeostasis and survival, which include protein folding, lipid biosynthesis, and calcium and redox
homeostasis (Rutkowski & Kaufman, 2004). However, diseases arise when the normal function of the
organelle ceases due to errors like misfoldings of proteins, oversynthesis of molecules, etc.
Understanding the mechanism on how the organelle manages to survive and counteract with these
abnormalities is a big deal to know the nature of related diseases. This can be started by investigating the
features on how Unfolding Protein Response (UPR) works which is triggered due to ER stress.
Therefore, the factors and effects which cause either the normal or overactivation of the response will be
studied and compared. Moreover, according to Ozcan & Tabas (2012) during the past decade, research
on how cells can sense and cope with ER stress has rapidly progressed. There is also compelling
ongoing research that suggests one or more branches of the UPR are important in the pathogenesis of
several diseases. However, many unanswered questions remain. They also added that a detailed
understanding of the mechanisms and consequences involved in these processes will be important for
translating our knowledge into novel therapeutic approaches.

The research of Marton et al. (2017) weighs on the two fates of the cell through the factors
affecting the UPR. In this study, they focused on the kinetical features of PERK branch of UPR and the
contribution of IRE-1 and PERK arms in life-and-death decision. Progress in the field has provided insight
into the regulatory mechanisms and signalling crosstalk of the three branches of the UPR, which are
initiated by these stress sensors protein kinase RNA-like ER kinase (PERK), inositol-requiring protein 1α
(IRE1α) and the other one is activating transcription factor 6 (ATF6) (Hetz, 2012). This study connects
and correlates with the two other studies because they are its applications to the medical field. The
interplay of these stress sensors is very crucial to determine the nature of diseases due to ER stress and
develop possible approach for therapeutic techniques to alleviate these diseases using UPR fluctuations.

“Protein misfolding in the endoplasmic reticulum as a conduit to human disease” by Wang &
Kaufman (2016) further explains what will happen if the cell stops to function and proceed to apoptosis
due to UPR abnormalities. They also noted that protein folding is the most error-prone step in gene
expression. The ER has essential roles in physiologic regulation of many processes. Accumulating
evidence indicates that pathologic conditions that interfere with ER homeostasis give rise to chronic
activation of the UPR, which contributes to the pathogenesis of many diseases (Ozcan & Tabas, 2012).
These include neurodegenerative disorders, type 2 diabetes, atherosclerosis, liver disease, and cancer.
On the other hand in terms reproductive pathology, mounting evidence shows that ER stress and
associated UPR signaling are important contributors to the normal functioning of reproductive tissues,
including endometrial menstrual cycle changes, regulation of gametogenesis, development of the
preimplantation embryo and placenta as well as maintenance of pregnancy and initiation of labor but
diisruption of ER homeostasis as a result of excess accumulation of unfolded/misfolded proteins due to
prolonged and/or severe ER stress is involved in several pathologies that negatively impact on oogenesis
and spermatogenesis, and induce endometriosis and endometrial/ovarian cancers as well as various
pregnancy complications that result in preeclampsia, Fetal growth restriction, and preterm birth (Guzel et
al., 2017) In the case of cancer, many studies have demonstrated a crucial role for the UPR in both
tumour cells and stromal cells in the tumour microenvironment as cancer develops and progresses. The
question “to stimulate or inhibit” UPR to increase cancer cell sensitivity to death is still a challenge since a
deeper knowledge of each tumor must be taken into account, to show the path and it is important to
consider that there are still no specific and clinically available “modulators,” either positive or negative, of
UPR (Corrazari, 2017).
Figure 1. Endoplasmic Reticulum (ER) homeostasis/stress and the unfolded protein response (UPR) signaling in
physiopathologic conditions. (A) In physiological (unstressed) states, these transducers bind to the folding chaperone
GRP78 and keep the ER quiescent (B) ER stress inducers accumulate unfolded/misfolded proteins in the ER lumen by
impairing protein folding. Higher GRP78 affinity for unfolded/misfolded proteins dissociates GRP78 from ATF6, PERK
and IRE1, enabling GRP78 unfolded/misfolded protein binding that then initiates three UPR signaling cascades (Guzel
et al., 2017).

On the other hand, “Potential for therapeutic manipulation of the UPR in disease” by Park &
Ozcan (2013) focuses on the molecular signaling pathways of the UPR and suggest possible ways to
target this response for therapeutic purposes. The primary role of the UPR is to maintain and re-establish
ER homeostasis. Moreover, under physiological conditions, the UPR is activated as a cellular survival
program that protects the cell from ER stress and helps it recover from damage or increased work
overloads. In essence, the UPR functions to reset the ER, from the “stress” state back to its normal
condition (Marciniak, 2006). The study highlighted UPR elements in pathophysiology and studied these
elements to point their significance in affecting the UPR activation for the diseases: Obesity and type 2
diabetes, Wolcott-Rallison syndrome, Cystic fibrosis, Neurodegenerative diseases and Cancer. Park &
Ozcan (2013) emphasized that there is no universal solution that is relevant for developing strategies to
manipulate ER stress in different diseases and effective solutions will have to be individualized for each
condition. Moreover, Ozcan & Tabas (2012) added that although relieving ER stress may be beneficial in
certain diseases, such as T2D, atherosclerosis, and neurodegeneration, increasing ER stress may be
helpful in other disease processes, such as cancer and possibly certain types of viral infections.
Therefore, control of the levels of UPR activation and its elements is essential for the treatment of
diseases. Aside from this, it was found out that chemical chaperones are small molecules that prevent
protein aggregation, facilitate protein folding and reduce ER stress — both in vitro and in vivo — by
stabilizing protein-folding intermediates. The chemical chaperone tauroursodeoxycholic acid has been
studied intensively as a treatment for many diseases. Its therapeutic potential has been demonstrated in
mouse models of diabetes (Engin, 2013).

Figure 2. ER stress and UPR pathways as therapeutic targets

(Wang, 2015).

Through the studies of Wang & Kaufman (2016) and Park & Ozcan (2013) it was found out that
UPR activation were different for every disease. The onset of diseases varies due to the differences of
participation of UPR elements, stress sensors, branches, etc. affecting the activation of UPR and state of
ER stress. Therefore, solutions and therapeutic aids for these are not universal and every related disease
has its therapeutic technique to prevent its adverse effects. Another constraint is that the studies of UPR
activation and ER stress was not well-developed enough for some diseases. It was even point out by the
recent study by Corazzi, et al. (2017) “Endoplasmic Reticulum Stress, Unfolded Protein Response, and
Cancer Cell Fate” that an intensive effort is still required to (i) better define the role of UPR in specific
tumors, (ii) unveil the role of each UPR branches in each tumor, and (iii) identify/develop drugs with high
target specificity and low side effects. However, as stipulated again by Ozcan & Tabas (2012), during the
past decade, research on how cells can sense and cope with ER stress has rapidly progressed.

Summary

Endoplasmic reticulum, specifically the rough ER functions as a protein factory and as well as
quality control of protein being synthesized. It also plays a role in maintaining protein folding homeostasis,
thereby controlling ER stress of the cell. ER stress is caused by misfoldings of the protein, thereby causes
diverse types of ailment. The cell alleviates this stress by the process called unfolding protein response
that reset the ER, from the “stress” state back to its normal condition (Marciniak, 2006). UPR has three
branches which is responsible for “resetting” the cell back to its normal condition: PERK, IRE1, and ATF6.
PERK and IRE1 are responsible in the mechanism and signaling cross-talk of the three branches, and
this two UPR elements also dictates whether a cell will undergo autophagy or apoptosis. Pathologic
conditions that interfere with ER homeostasis give rise to chronic activation of the UPR, which contributes
to the pathogenesis of many diseases. The onset of diseases varies due to the differences of participation
of UPR elements, stress sensors, branches, etc. affecting the activation of UPR and state of ER stress.
Therefore, solutions and therapeutic aids for these are not universal and every related disease has its
therapeutic technique to prevent its adverse effects.

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