1.

Name: Shiza Malik

2. Registration Number: Nust
3. Discipline: Applied Biosciences.
4. Institute: Atta-Ur-Rehman School Of Applied Biosciences
5. Supervisor: Dr. Rumeza Hanif

Topic: Effect of CXCR4 in Breast Cancer Signaling Pathways

Introduction:

1 the Uncontrolled division of the cells ‘more specifically called

Breast cancer is associated with
as the tumor formation’ within the breast region, these tumors may be early stage begnin cancer
tumors which makes the cancer curable but if The tumor detaches itself from the breast region
and enters the blood stream and lymphatic vessels. It will become cancerous and will metastasis
other distinct parts of the body like liver kidney, etc. in such cases even if the tumor is removed
surgically or by radiation, chances of remission will be fairly significant these malignant tumor
could be seen through x-rays and can be observed as lumps on the breast.1

Breast cancer can be divided into different types depending of the type of tissues being invaded
including Ductal carcinoma in situ (DCIS), Invasive (or infiltrating) ductal carcinoma
(IDC) and Invasive Lobular Carcinoma (ILC).Some other less common types of breast
cancer are: Inflammatory breast cancer, Paget disease, Phyllodes tumor and
Angiosarcome.11

Breast cancer is the second most prevalent cancer in women after various skin cancers effecting
one in every eight women worldwide. (American Cancer Society, 2002) World Health
Organization estimated that each year 182,000 women’s are affected with breast cancer
(American Cancer Society, 2002) .On international basis the breast cancer is not only rising in
eastern states and European countries but now also becoming more common in Asian countries
like japan, china, Pakistan3. Breast cancer is a complex disease which involves several
interlinking signalling pathways that play an important role in the development of complex
regulatory network of genes involved in breast cancer 8.

Incidences in Pakistan
Approximately 1 in every 9 women in Pakistan is likely to be diagnosed with breast cancer
.The incidence rate of breast cancer is increased among all Asian countries after Israel and
almost 2.5 times higher than ten neighbouring countries like Iran and India. It is accounting for
34.6% of female breast cancer in young age which negatively effects the prognosis 5, Pakistan
has a strong genetic support for breast cancer development that leads to increased risk through
cousins marriages 6.
Causes of Breast Cancer 11
There are many risk factors that can be regarded as responsible for breast cancer development
some of them are age , family history with breast cancer , genetics mutations, races , altered
hormonal level menstrual cycle , dense breast and some reproductive factors related to
pregnancy , breast feeding , fertility drugs intake or hormonal therapies and hormonal birth
control pills as well as some behavioural factors are also involved sometimes like obesity,
smoking or alcohol intake , overall inactive lifestyle , and environmental factors that may
encourage breast cancer development.

Treatments of Breast Cancer 11

Several different treatment options are available for breast cancer, such as:
surgery, chemotherapy, hormonal treatment, biological treatment for immunity booster,
and radiation therapies .and some specific targeted therapies that work by blocking specific
breast cancer promotor genes like cyclin-dependent kinase CDK4 and CDK6 and therapy
against HER 2, and against mTOR, a protein that promotes cell growth and division.
2
Problem Statement:

The conventional therapies for breast cancer like surgery, hormonal treatments, radiation and
chemotherapy are extremely expensive and chances of remission remains fairly significant
making extremely less chances of survival .Moreover, these therapies are highly unspecific and
non-targeted. So, the need of a more targeted therapy arises for breast cancer arises. 11
The role of these chemokines in breast cancer had already been studied through various
researches that show that these chemokines effect the progression of breast cancer and the
effects of blocking or enhancing these chemokines on breast cancer regression and
development respectively13. Previous studies shows that chemokines play an important
inhibitory targets to control the effect of metastasis in BC this can be inferred by the High
levels of CXCR4 that results in increased expression of various genes/proteins involved in
signalling pathway such as PI3K, AKT, HIF-1 alpha and ER alpha that leads the tumor towards
metastasis.14
CXC chemokine ligand 12 (CXCL12), previously designated stromal cell-derived factor 1
(SDF-1) is a small chemotactic cytokine belonging to the CXC chemokine family and it is
constitutively expressed in various organs (Muller et al. 2001). It was first cloned from a bone
marrow-derived stromal cell line and was later identified as a pre-B-cell growth stimulating
factor (Tashiro et al., 1993). It is a ligand of CXCR4 which is a G-protein coupled receptor to
promote intravasation of breast cancer cells from the primary tumor into blood vessels and
extravasation at sites of metastases (Fernandis et al., 2004). CXCR4 is also positively and
directly correlated with HER-2.14 Up regulated expression of CXCR4 in breast cancer cells is
a key determinant of tumor progression.
Materials and methods

We will use the kinetic logic formalism of Biological Regulatory Network to find the
complexity of interactions among genes and proteins involved in signaling network related to
BC. We will use the computational tree logic (CTL) formalism for detecting suitable
parameters which will be checked through model checkers including SMBioNet software. By
selecting the different parameters according to the biological observations and then applying
them in software GENOTECH. We will generate an asynchronous state graph, by utilizing the
BRN system to analyse the expression levels of genes and proteins involved in signalling
networks of breast cancer.
These in-silico studies will be followed by the wet lab experimentations where we will check
the expression of various genes involved in breast cancer through RT-PCR technique

Aims and objectives

1. To study the biological regulatory network involved in BC metastasis
2. To identify the role of chemokines such as CXCL12 and CXCR4 in BRN.
3. To examine the complex3 dynamical interaction among genes and proteins associated to
BC by using computational modelling formalism
4. To understand that specific CXCR4 inhibition will impaired the proliferation and
migration potential of metastatic cells thus establishing that CXCR4 could be a general
and attractive target for anticancer
5. To examine differences in expression pattern of different signaling networks in breast
cancer patients as compared to healthy individuals.
6. To find co-relation between the expression level of CXCL12 and other genetic
regulations computationally to find their effecting rate of metastasis in breast cancer.

Hypothesis:
Our hypotheses is to formulate a conceivable and more convenient approach towards
developing a conclusive targeted drug for curing breast cancer using a computational analytic
approach that will help to elaborate the distinctive role of CXC12 and CXCR4 that regulate
HER-2 and CCL17 by controlling the dynamic network systems of signalling pathways
involving PI3K, AKT, HIF1 alpha, P53, ER genes that play a significant role in breast cancer
development.

Timeline:

FEBRAURY MARCH APRIL MAY JUNE

Literature Modelling using Wet lab Result Thesis defence
review and SMBioNet experimentations validation and
modelling using software compilation
GENOTECH
software
 REFERENCES

1 .American Cancer Society. What is Breast Cancer? Medical Review: 09/25/2014. Revised: May 4, 2016.

2. Howlader N, Noone AM, Krapcho M, et al. SEER Cancer Statistics Review, 1975-2012. http://seer.cancer.gov/csr/1975_2012/, based on

November 2014 SEER data submission, posted to the SEER web site, April 2015. Bethesda, MD: National Cancer Institute, 2015.

2. Copeland G, Lake A, Firth R, et al. Cancer in North America: 2008-2012. Volume One: Combined Cancer Incidence for the United States,

Canada and North America. Springfield, IL: North American Association of Central Cancer Registries, Inc, June 2015.

3 http://emedicine.medscape.com/article/1697353-overview

4 http://www.cancer.org/cancer/breastcancer/detailedguide/breast-cancer-breast-cancer-types
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5 Sohail S, Alam SN. Breast cancer in Pakistan: awareness and early detection. J Coll Physicians Surg Pak 2007; 17:711-2.

6 https://www.ncbi.nlm.nih.gov/pubmed/24289553

7 Centers for Disease Control. Risk Factors for Young Women. Reviewed: March 13, 2014 Updated: March 13, 2014.

8 https://peerj.com/articles/2542/

9 Foster C, Evans DGR, Eeles R, Eccles D, Ashley S, Brooks L, et al: Predictive testing for BRCA1/2: attributes, risk perception and

management in a multi-centre clinical cohort. Br J Cancer. 2002, 86: 1209-1216. 10.1038/sj.bjc.6600253

10 Foster C, Watson M, Moynihan C, Ardern-Jones A, Eeles R: Genetic testing for breast and ovarian cancer predisposition: cancer burden

and responsibility. J Health Psychol. 2002, 7: 469-484. 10.1177/1359105302007004627

11 http://www.cancer.org/acs/groups/content/@research/documents/document/acspc-046381.pdf

12 http://onlinelibrary.wiley.com/doi/10.1111/j.1349-7006.2006.00307.x/pdf

13 https://www.hindawi.com/journals/jir/2014/849720/

14 https://www.ncbi.nlm.nih.gov/pubmed/21479551
(Student’s signature) (Supervisor’s signature)
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