GENETIC ENGINEERING AS AN EFFECTIVE

CANCER THERAPY: A FOCUS ON CRISPR

BY: SAI SRUTHI MOPURI

 OVERVIEW OF THE PRESENTATION

Project abstract What is cancer? What is Significance and

Why is it CRISPR? Its Putative targets Results, findings future research
and research for 3 cancers of
question important to brief history and and conclusion ideas
research? applications? focus
PROJECT ABSTRACT AND RESEARCH QUESTION
GENETIC ENGINEERING AND ITS THERAPEUTIC APPLICATION
Genetic Engineering is the direct manipulation of an organism’s
genetic material
• Single nucleotide shifts
• Exogenous gene expression

Why?

• Precision
• Accuracy
• Potential to correct genes causing lethal diseases

Research question?

• Using CRISPR Cas9 as genetic engineering tool for cancer pathologies

• Find CRISPR Cas9 targets for triple negative breast cancer, acute myeloid leukemia and
glioblastoma multiform
• CRISPR PCR for molecular profiling tumors
WHAT IS CANCER AND WHY IS IT IMPORTANT TO
RESEARCH?
 CANCER
 A group of diseases.

 Characterized by unregulated cell growth and the invasion and spread of cells from the site of origin (primary site) to
other sites in the body.

 Types(based on the location of primary) :

1) Carcinoma

2) Adenocarcinoma

3) Sarcoma

 Hallmarks of cancer

 Steps of cancer progression

 Cause: underlying mutations

Pancreatic cancer metastases tumor histology

SOURCE: Research gate publication 329762714
 HALLMARKS OF CANCER
Defined as the underlying properties that govern a cancerous cell.
Proposed by Hanahan and Weinberg in 2000.
6 defined hallmarks common to most cancers.

Source: Molecular Biology of Cancer Mechanisms Third Edition (1.1)

 WHY IS CANCER RESEARCH IMPORTANT?

One of the most lethal diseases and statistics suggest prevalence has
increased linearly over time.
In 2018, there were 18.1 million new cases and 9.5 million cancer-related
deaths worldwide.
By 2040, the number of new cancer cases per year is expected to rise to 29.5
million with the number of cancer-related deaths to 16.4 million. 
Cancer mortality rate : 158.3 per 100,000 men and women per year
Approximately there is a 39.5% probability an individual will be diagnosed
with cancer at some point during their lifetimes.
WHAT IS CRISPR AND ITS APPLICATIONS?
 CRISPR-CAS9

A technique that allows for the highly specific and rapid modification of DNA in a
genome.
Significance:
• Versatile technique and multiple allele correction.
• Lesser duration- reducing time span for gene editing from months or years to days.
• Not species-specific.
• High degree of flexibility and accuracy in cutting and pasting DNA.
 Hailed as “the” technology of the decade by the Nature Journal.
 CRISPR CAS9 MECHANISM

Source:CRISPR/Cas9 - CRISPR Biotech Engineering (crispr-bte.com)

CRISPR AND CANCER
Putative targets for 3 cancers of focus
 TRIPLE NEGATIVE BREAST CANCER (TNBC)

 Constitutes 10–20% of all breast tumors

 Characterized by a lack of expression of estrogen receptor (ER), progesterone receptor (PR)
and HER2.
 Generally high grade, aggressive tumors.
 High rate of distant metastasis
 Poorer disease-specific survival than other breast cancer subtypes.
 Calls for novel therapeutic strategies to improve patient care and survival.
 Associated with a shorter median time of relapse and death.
CRISPR TARGETS FOR TNBC
 PUTATIVE TARGETS FOR GLIOBLASTOMA
 Glioblastoma Multiforme (GBM) is an aggressive variant of brain cancer.
 Site of primary:- astrocyte cells of the brain.
  Form in the cerebral white matter, grow quickly, and can become very large before
producing symptoms, which contributes to its fast, uncontrolled metastasis.

Tumor histology of GBM

 CRISPR TARGETS
PUTATIVE TARGETS FOR GLIOBLASTOMA MULTIFORME

TRP53/P53

PD-1/PD-L1

NF-1

TERT

PTEN

MAP4K4

Epidermal growth factor receptor variant III (EGFRvIII)

 PUTATIVE TARGETS FOR ACUTE MYELOID LEUKEMIA
 Acute myeloid leukemia (AML) is the most common leukemia among the adult population and accounts for about
80% of all cases.
 It is characterized by clonal expansion of immature "blast cells" in the peripheral blood and bone marrow.
 AML is characterized by mutations of the genes involved in hematopoiesis (blood cell production).
 AML is a highly heterogeneous disease with a variable prognosis. It can result from genetic mutations,
chromosomal translocations, or changes in molecular levels.

FIG:-1 PEDIATRIC AML PATHOPHYSIOLOGY

 TARGETS FOR AML
PUTATIVE TARGETS FOR ACUTE MYELOID LEUKEMIA

DNMT3A WNT–β-catenin amplification(INCREASES IMMUNE

SURVELLIENCE)

CRM1 inhibitors IDH inhibitors

FLT3 inhibitors EZH2

Monoclonal antibody CD33 RUNX1

Induction of IDH2 PD-1 (Common target for multiple cancers)

R140Q Mutation

DOT1L inhibitor Pinometostat (EPZ-5676) EZH2 inhibitor (DZNep (3-deoxyadenosine) 

 SIGNIFICANCE AND TAKEAWAYS
 Cancer therapies need urgent modifications to promote larger survival rates.
 Gene therapies confer to better survival.
 CRISPR is the most efficient gene editing technique for the three cancer types studied.
 Compiled and studied putative targets for three aggressive cancers.
 Putative targets of my research may be further studied and engineered to use as gene
therapy.
 SUMMARY OF MY PRESENTATION
 Began with a brief introduction to cancer.
 Understood the characteristics and mechanisms of cancer formation.
 Understood about CRISPR Cas9 technology and its applications in cancer
pathologies.
 Understood putative targets for CRISPR for Triple negative breast cancer, Acute
Myeloid Leukemia and Glioblastoma Multiforme.
 Understood the importance of CRISPR for cancer and significance of my research.
THANK YOU!
AND A HUGE THANK YOU TO MY MENTOR DR. JOSE DELGADO