Major Credit SeminarOn

Advancements in Cancer Research with Special Reference to Pathogenesis and Diagnosis

Presented by

Dr. Rahul G. KadamPh.D Scholar

Roll NO. P1661

Present Cancer Scenario.. • Out of the total deaths in world 12-13 % deaths are due to cancer..

• In world 8.2 million people died due to cancer during 2013.

• By 2030 deaths are expected to rise up to 12.0 million.

• The cancer deaths are exceeding the cardiac deaths.

• Every day 2000 peoples died by cancer worldwide.

• Among all these losses

40 % deaths can be avoided

by early diagnosis and according

treatment.

“ An abnormal mass of tissue, the growth of which exceeds and is uncoordinated with that of the normal tissues and persists in the same excessive manner even after cessation of the stimuli that evoked the change.”

New growth composed of cells originally derived from normal tissues, that have undergone heritable genetic changes allowing them to become relatively unresponsive to normal growth controls and to expand beyond their normal anatomical boundaries.

Cancer

TYPES OF NEOPLASM

BENIGN NEOPLASMS

Designated by the suffix – ‘oma’

• Adenoma – glandular epithelium

• Papilloma – epithelial tumor forming finger like projections with a connective tissue core

MALIGNANT NEOPLASMS

• Sarcoma – from mesenchymal tissue

• Carcinoma – from epithelial tissue

CLASSIFICATION/NOMENCLATURE

HISTOGENIC CLASSIFICATION

Based on the origin of neoplastic cell types.:

• Mesenchymal tumours: Arise in cells of embryonic mesoderm

Benign – oma; malignant – sarcoma

• Epithelial tumours:

Glandular – adenoma/adenocarcinoma

Squamous / basal

• Non epithelial: fibrous, cartilage, bone, muscle, etc

Some tumors have more than one parenchymal cell type

– Mixed tumors –derived from a single germ cell layer that differentiates into more than one cell type.

– Teratomas – made of a variety of parenchymal cell types that derive from more than one germ cell layer formed by totipotent cells that are able to form ectoderm, endoderm & mesoderm.

Tissue of origin Benign Malignant

A)Tumors of mesenchymal origin

1.Connective tissue and derivatives

Fibroma

Lipoma

Chondroma

Osteoma

myxoma

Fibrosarcoma

Liposarcoma

Chondrosarcoma

Osteogenic sarcoma

myxosarcoma

2.Endothelial and related tissues

Hemangioma

Lymphangioma

Meningioma

-

Hemangiosarcoma

Lymphangiosarcoma

Invasive meningioma

mesothelioma

3.Tumours of Hematopoietic cells

-

Lymphoma

-

-

Lymphoid leukaemia

Lymphosarcoma

Myeloid leukaemia

Multiple myeloma

4.Tumours of Muscle Leiomyoma

Rhabdomyoma

Leiomyosarcoma

Rhabdomyosarcoma

B)Tumors of Epithelial origin Papilloma

-

Adenoma

melanoma

Transititonal cell papilloma

-

Squamous cell carcinoma

Basal cell carcinoma

Adenocarcinoma

Melano carcinoma

Transitional cell carcinoma

Seminoma

c)Tumours of nervous tissue

neuroma neuroblastomaWHO & IARC ,Lyon, 2000

DEVELOPMENT OF

CANCER

METALOPROTINEASEMETALOPROTINEASE

Development of malignant tumour

Less than 10 mutation required

Etiology of cancerEtiology of cancer

Intrinsic factors • HEREDITY• AGE• PIGMENTATION . SEX . TUMOUR IMMUNITY

EXTRINSIC FACTOR •CHEMICALS•RADIANT ENERGY•CHRONIC IRRITATION•HORMONE•PARASITES•ONCOGENIC VIRUS

Chronic irritation : Kangari,Parasite :Gongylonema.neoplasticum

Environmental Carcinogens

• A cancer-causing agent• Three main types:

– Chemical– Physical (radiation)– Biological (especially virus)

• Direct-acting• Indirect-acting (must be metabolized to activated

metabolic forms • Direct-acting carcinogens are already electrophilic• Indirect-acting carcinogens are metabolically activated

into electrophilic species

How Cancer Arises

1. Cancer cells violate the civic rules that govern normal cells by not responding to go-signals for proliferation and stop-signals for reproduction

2.  Cancer cells descend from a common ancestral cell:

clonal origin. But at some point one of the off-springs mutate that becomes worse with more mutation, and finally the accumulated mutated cells disobey all civic controls of normal cells in a tissue, becoming invasive and malign.

3. Since mutations occur at the gene level, that is, DNA molecules that reside in the nuclei of the cells, most human cancer be traced there.

Molecular Basis of Cancer

• Tumors are monoclonal proliferation

• Tumors carry genetic defect that are not lethal (inherited or acquired)

• Transformed cells acquire gene defects that allow them to form tumor

What are the genes involved in Cancer?

• A lot of genes!• Like what?• Genes promote growth eg. RAS• Genes inhibit growth eg. P53• Genes control apoptosis eg.

Bcl-2• Genes of DNA repair

• And others….

Functional classification

Up regulated genes (%)

Down regulated genes(%)

Metabolism 21 30

Cell adhesion 4 4

Cell structure 3 6

Immune response

6 9

Functional classification of genes discriminating the normal rat

Mammary gland from NMU-induced tumors

( Marsen M.et al., 2005)

Oncogenes and tumor suppressors

Current gene list and technology

Knudsen’s “two- hit” hypothesis

Two separate mutations, one in each allele of a tumorsuppressor gene, are needed to cause cancer

18

How cellular oncogenes arise

Cellular oncogenes arise from proto-oncogenes

Proto-oncogenes are not bad genes.

Normal genes for regulation of cell proliferation and survivalWhen it change the structure and activity by mutation: causing cancerGain-of-function mutation

• Targeted genes:– 1. Proto-oncogenes (oncogenes)– 2. Tumor suppressor genes– 3. Genes controlling apoptosis– 4. Genes regulating DNA repair

• Other genes involved:– Genes regulating angiogenesis– Genes enhancing invasion and metastasis

• Carcinogenesis is a multistep process– At both genetic and phenotypic levels– Progression results from accumulation of genetic

defects

The cancer –related gene produce six basic change.

What does a cell need to be “cancer”?

The hallmark of cancer

1. ONCOPROTEIN2. RB GENE3. BAX gene4. TELOMERASE5. VEGE, bFGC,

THROMBOSPODIN-1 (Sheibani et al.,1999)

6. E-CADHERINS , BETA CATEINS

Most cells are quiescent and are in the G0 (inactive) part of the cell cycle

Most adult cells are NOT actively dividing

Mutation in one of the four genes that regulate cell cycle. RB.CDK4,CyclinD ,CDKN2A

G1

S

RB

Adenovirus E1AHPV E7

SV40 Lg T

APOPTOSIS

p53

Adenovirus E1B(55K)HPV E6

SV40 Lg TAdenovirus E1B (19K)(Bcl2-like)

Viral Oncogenes Induce Proliferation and Suppress Apoptosis

RB protein unable to bind the E2F

DNA viruses can ruin two of the best understood Tumour supressor gene.

P53 gene is a monitor of stress.

Molecular basis of cancer

Classification of oncogenes

1. Growth factors – c sis,

2. Growth factor receptors (RTK) - ERBB2, fms

3. Non receptor tyrosine kinases - abl, src

4. GTP binding - RAS

5. DNA damage repair - ATM, MSH2, B cl2

6. Serine/ threonine kinases

7.Nuclear Transcription Factors – MYC,MYB,JUN, FOS

8. Misc - cell surface APC/ DCC

Imogene : Gene mutated such that the protein is produced in higher quantity or is more active and initiates tumor formationImogene : Gene mutated such that the protein is produced in higher quantity or is more active and initiates tumor formation

How Cellular Oncogenes ArisePoint mutation

.

Tumor suppressor: Gene that normally inhibits tumor formation; mutation or poor expression releases the inhibition

MDM2 ,triggers transcription for many gene

Key tumor suppressor genes are mutated

Molecular Policeman

 BRCA1 and BRCA2, two famous genes whose mutations confer a high increased risk of breast cancer on carriers, are both associated with a large number of DNA repair pathways, 

XERODERMA PIGMENTATION

Early detection is the key!

Cancer diagnosis comprises of

.involves evaluation of the patient’s history,

clinical examinations

paraneoplastic disorder

review of laboratory test results

radiological data, (X-ray.CT scan ,MRI. Ultrasound imaging)

Biopsy

Cancer staging

Molecular marker.(PCR and RT PCR based technique.)

Diagnostic approaches of tumor.

Stage 0: carcinoma in situ.

Stage I: cancers are localized to one part of the body

Stage II: cancers are locally advanced.

Stage :Whether a cancer is designated as Stage II or Stage III can depend on the specific type of cancer; 

Stage IV: cancers have often metastasized, or spread to other organs or throughout the body.

TNM SYSTEM COMMONLY USED METHODOF STAGING BASD ON PRIMARY LESION. .

Grade  

GX Grade cannot be assessed (Undetermined grade)

G1 Well-differentiated (Low grade)

G2 Moderately differentiated (Intermediate grade)

G3 Poorly differentiated (High grade)

G4 Undifferentiated (High grade)

The American Joint Committee on Cancer recommends the following

guidelines for grading tumors (1):

IMAGING

Malignancy is based on imaging information ,later confirmed on

histology

Ultrasound (kondyo et al.,2008)

Computed topography,(Drosot et al.,1996)

Magnetic resonance imaging(MRI)(kaiser et al 1992)

Metabolic and functional ImagingMolecular imaging with magnetic resonance spectroscopy.(MRS)

Position emission topography,(PET)(Grahek D et al 2004)

Cytology and Histopathological Technique

1. Still a gold standard for diagnosis of tumour.

Recent technique ImagingIHCPCRFlow cytometary FISHMICROARRAYNANOTECHNOLOGY

• What is tumor marker? Tumor markers are glycoproteins produced by tumor cells or by other body cells in response to cancer or other conditions.

As tumor cells multiply, spreads & tissue is damaged TMs increase in concentration.

• How it produced?

Tumor Markers

• Where do the TMs found? These substances can be found

in the blood (plasma, serum), urine, saliva, tissue fluid, in the tumor tissue or in other tissues.

Sr. Sr. No.No. Disease/TissueDisease/Tissue MarkersMarkers

11 Bladder cancerBladder cancer BTA, NMP22, CEA, CA125, CA19-9BTA, NMP22, CEA, CA125, CA19-9

22 Breast cancerBreast cancer CA15-3, CEA, CA27-29CA15-3, CEA, CA27-29

33 Colorectal cancerColorectal cancer CEA, CA 19-9CEA, CA 19-9

44 GestationalGestational HCGHCG

55 Liver cancerLiver cancer AFPAFP

66 Lung CancerLung Cancer CEA, NSECEA, NSE

77 Melanoma sink Melanoma sink cancercancer

TA-90, S100TA-90, S100

88 Multiple myelomaMultiple myeloma B2MB2M

99 Ovarian CancerOvarian Cancer CA125, CA72-4, HCG, AFPCA125, CA72-4, HCG, AFP

1010 Pancreatic CancerPancreatic Cancer CA 19-9, CEACA 19-9, CEA

1111 Prostate CancerProstate Cancer PSA, PAPPSA, PAP

1212 Gastric CancerGastric Cancer CEACEA

1313 Testicular CancerTesticular Cancer HCG, AFP, PAPHCG, AFP, PAP

List of commonly used TMs

• Ag –Ab based Techniques :

i) ELISA

ii) Radio-immunoassay

iii) Flow Cytometry

iv) Immunohistochemistry

• Molecular Genetic Techniques :

i) PCR / RT PCR

ii) Fluorescence in situ Hybridization ((FISH)

iii) Comparative Genomic Hybridization (CGH)

• Other Techniques :

i) Spectophotometry

Tumor Markers detection Methods

Immuno-histochemistry has been utilized extensively to determine estrogen, progesterone and Her-2 neu receptor status in breast cancer in predicting response to therapy (Diaz, Leslie et al.,2005)

Detection of over expression of c-erbB2 oncoproteins by ICH in canine mammary tumour.( Mayilkumar K et al ,.2009)

Immuno- histochemistry(IHC)

IHC based on specific antigenic determinants in the cell of tissue by use of polyclonal or monoclonal antibodies.(Ramos-Vara et al., 2005)

Immunohistochemistry (IHC)

• Ag + [Ab + fluorescent dye]

• Detection of protein of tumors

by using specific antisera &

MCAb directed against them.

• Detection of surface receptors

to intracellular matrix to hormone

can be detected with ease.

• Help ti determine the primary

site of metastatic tumor.(Ramos-Vara et al., 2005)

A single gene chip can even hold representative fragments from the entire human genome.

Different microrray labeling kit

• Present in perfectly normal conditions

• Act as chaperones making sure that the cell’s proteins are in the right shape and in the right place at the right time.

• HSPs also help to shuttle proteins • Transport old proteins to

“garbage disposals” /proteasome inside the cell.

Heat shock Protein

• For normal function of p53 it require transient interaction of Hsp 90 and then degraded

• Mutation of p53 • Mutated p53 have

unstable conformation

• Extended interaction with Hsp 90 and prevent their normal degradation

• Accumulation of mutant p53

• Over-expression of HSP70 and HSP90 correlates with a bad prognosis of tumor

• Over expression of HSP27 in leukemia osteosarcoma ovarian cancer, prostate cancer

• Overexpression of HSP70 indicates high grade malignancy

According to the US National Cancer Institute (OTIR, 2006) “Nanotechnology willchange the very foundations of cancer diagnosis, treatment, and prevention”

NANOTECHNOLOGY IN CANCER

. Nanotechnology will make possible to run test without physically altering the cells or tissue

Cancer nanotechnology is emerging as a new field of interdisciplinary research, cutting across the disciplines of biology, chemistry, engineering, and medicine, and is expected to lead to major advances in cancer detection, diagnosis, and treatment .(Menon U, Jacobs IJ. 2000, Ferrari M. 2005.)

Schematic diagram showing nanotechnology applications in cancer through molecular tumor imaging, early detection, molecular diagnosis, targeted therapy, and cancer bioinformatics

Nanodevices Are Small Enough to Enter Cells

Cell

White blood cell

Water molecule

Nanodevices

Nanodevices

Nanodevices Can Improve Cancer Detection and Diagnosis

ImagingNanotechnology Physical Exam,Symptoms

Nanodevices Can Improve Sensitivity

and determinewhich cells arecancerous orprecancerous.

Precancerous cells

Normal cells

Nanodevices could potentiallyenter cells

Precancerous cells

Normal cells

Nanodevices Can Preserve Patients’ Samples

Cells from patientCells preserved

Active state preserved

Cells altered Active state lost

Additional tests

Cells from patient

Nanotechnology Tests

Traditional Tests

Nanodevices Can Make Cancer TestsFaster and More Efficient

Patient A Patient B

Laser Capture Microdissection

Cancer prevention

Conclusion

1.Histopathology remains the standard conventional method.

2. Recent technique such as imaging ,ICH ,PCR ,flow cytometry ,FISH, CSH ,and microarry nanotechnalogy contribute a major break through in diagnosis prognosis of cancer.

3.In future a multimodal imaging approach will evolve, enhancing diagnostic accuracy thus minimizing loss of lives due to cancer.

Cosmic Energy essential to maintain the order of our life and expand our consciousness, it is base for our all action and functions and to lead healthy and happy life. To remove all disorder of body. More cosmic energy is obtained through MEDITATION.