Concepts and Methodology in Cancer Diagnostics

An Immunological, Biochemical and Molecular Approach

1st Edition - November 5, 2024
Imprint: Academic Press
Authors: Seema Nayak, Anupam Kumar Sriwastava
Language: English
Paperback ISBN:
9 7 8 - 0 - 4 4 3 - 2 3 5 9 3 - 1
eBook ISBN:
9 7 8 - 0 - 4 4 3 - 2 3 5 9 4 - 8

Concepts and Methodology in Cancer Diagnostics: An Immunological, Biochemical and Molecular Approach provides an exclusive combination of all basics and recent technique… Read more

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Concepts and Methodology in Cancer Diagnostics: An Immunological, Biochemical and Molecular Approach provides an exclusive combination of all basics and recent techniques, methodology, and advancements in the field of cancer diagnostics. With a strong focus on the procedures and techniques involved in cancer diagnostics, content also gives insights into the molecular tests required for different tumors for the administration of targeted drugs and precision medicine. In 11 chapters. The book also introduces the methods and technologies based on DNA and RNA based biomarkers, biochemical and immunological markers, genetic signatures, epigenetic changes and exosomes in blood, and tissue and body fluids.

Readers will find a compilation of imperative topics and lists of the necessary materials and reagents needed, step-by-step instructions, readily reproducible laboratory protocols, as well as tips on troubleshooting and known pitfalls. This book is a valuable resource for medical professionals and researchers who want to understand the concepts of validated protocols and methodologies in diagnosing cancer, its prognosis, and predictive value.

Title of Book
Cover image
Title page
Table of Contents
Copyright
Dedication
Preface
Acknowledgments
Chapter One. Introduction to tumor diagnostic and prognostic: Differences between conventional and current approaches
Introduction
Conventional tumor diagnostic techniques
Other advance methods of cancer detection
Disadvantage of current cancer detecting methods
Future perspective
Chapter Two. Blood and other body fluid based biomarkers in tumor cells—Methods and protocol
Introduction
Tumor markers fall into one of several groups
Techniques: Most common methods for detection blood based biomarker in tumor cells are as mentioned below
Electrochemical sandwich ELISA
Molecular Techniques: Required DNA and RNA techniques
Quantitative polymerase chain reaction
Drawbacks
A successful RT-PCR assay for the detection of disseminated tumor cells in blood must meet several requirements, among which the following are the most important
Presenting some common blood-based biomarkers associated with tumor cells with their methods of identification including protocol
Advantages
Disadvatages
Procedure
Applications
Advantages
Challenges
Antibody-based immunoaffinity capture
Aptamer-based immunoaffinity capture
Advantages
Applications
Chapter Three. Immunological markers in different cancers: Laboratory testing protocol and pitfalls
Introduction
Programmed cell death protein 1 (PD-1) and Programmed Death-Ligand 1 (PD-L1)
Clinical significance
Cytotoxic T-Lymphocyte Antigen-4 (CTLA-4)
Tumor-infiltrating lymphocytes (TILs)
Human Epidermal Growth Factor Receptor 2 (HER2)
Epstein–Barr virus (EBV)
Microsatellite instability (MSI) and DNA mismatch repair (MMR) status
Immune infiltration signatures
Tumor Mutation Burden (TMB)
Interleukin-2 (IL-2) and Interferon-gamma (IFN-γ)
Human papillomavirus (HPV)
Some FDA approved immunological markers used in particular cancer and the laboratory tests used to detect them are given bellow
Laboratory testing protocols of different immunological markers
Chapter Four. Cancer biomarkers based on liquid biopsies: Update and challenges
Introduction
The potential of liquid biopsy-based early detection
Liquid biopsy applications
Challenges and future directions
Circulating RNA
Messenger RNA (mRNA)
MicroRNA (miRNA)
Long noncoding RNA (lncRNA)
Detection methods of circulating RNAs
Types of tumor-derived metabolites
Detection methods
Clinical applications
Challenges and future directions
Clinical applications
Methodology of estimation of liquid biopsy-based cancer biomarkers
Sample collection
Sample processing and standardization
Analysis techniques
Next-generation sequencing (NGS)
Polymerase chain reaction (PCR)
Digital PCR
Mass spectrometry (MS)
Immunohistochemistry (IHC)
Enzyme-linked immunosorbent assay (ELISA)
Single-cell analysis
Fluorescence in situ hybridization (FISH)
Circulating tumor cells (CTC) detection
Exosome analysis
Quality control
Data analysis and validation
Clinical correlation
Reporting and documentation
Ethical considerations
Iterative optimization
Challenges with current screening
Applications in various cancers
Clinical adoption
Research and technology advances
Challenges of CBLB
Sensitivity and specificity
Heterogeneity of tumors
Standardization and validation
Biomarker dynamics and stability
False positives and false negatives
Cost and accessibility
Ethical and regulatory considerations
Rare mutations
Normal tissue contamination
Limited biomarkers
Technical challenges
Interpatient and intrapatient variability
Integration into clinical practice
Clinical validation and regulatory approval
Future perspectives of liquid biopsy
Monitoring treatment response and resistance
Cancer screening programs
Multiomics approaches
Minimal residual disease (MRD) detection
Incorporation into patient stratification and clinical guidelines
Integration with artificial intelligence (AI)
Conclusion
Chapter Five. Oral salivary markers in different cancers: Current scenario in techniques and methodology
Introduction
Unique advantages of saliva as a diagnostic fluid
Microbiota
Methodologies for detecting salivary markers
Current scenario in salivary marker research
Collection techniques
Analytical techniques
Isolation of RNA from saliva
Other methods for RNA isolation
Primer designing
Components RT-PCR reaction volume (μL)
Future directions and innovations
Emerging technologies
Integration with artificial intelligence and machine learning
Multiomics approaches
Personalized medicine
Ethical and regulatory considerations
Global perspectives
Conclusion
Chapter six. Common childhood cancers: Diagnostic and prognosis markers
Introduction
Leukemia
Brain and central nervous system (CNS) tumors
Neuroblastoma
Wilms tumor
Lymphoma
Rhabdomyosarcoma (RMS)
Retinoblastoma
Bone cancer
Methods available for diagnosis
Genomic and molecular analysis
Cytogenetic analysis
Protein expression analysis
MicroRNA profiling
Epigenetic analysis
Histone modification analysis
Minimal residual disease (MRD) detection
Functional assays
Diagnosis
Diagnosis by other methods
Blood and laboratory tests
Imaging studies
Biopsy
Bone marrow aspiration and biopsy
Cerebrospinal fluid examination
Staging and risk assessment
Prognosis
Histology and pathological features
Genetic and molecular markers
Tumor size and stage
Response to treatment
Age at diagnosis
Tumor marker levels
Immune markers
Location of the tumor
Treatment modality
Minimal residual disease
Metastasis
Prognostic markers for childhood cancers
Future perspective
Chapter seven. Oncometabolites in tumor cells: Current technique and procedure
Oncometabolites in tumor cells: Current technique and procedure
What are oncometabolites
Chapter Eight. Immunohistochemistry based biomarkers in diagnosis of different types of tumor cells: Techniques, procedures and troubleshooting
Heat induced retrieval by buffer
Protocol #1: Citrate buffer antigen retrieval method
Protocol #2: EDTA buffer antigen retrieval protocol
Inactivation of endogenous peroxidase
Secondary antibody reaction
Option 1—Biotin/ExtrAvidin detection
ExtrAvidin reaction
Option 2—Enzyme-labelled secondary antibody—Polymer based kit
Substrate preparation
Development
Counterstaining
Assay required for revalidation: Following assay required
Chapter Nine. Currently available immunohistochemical marker for tumor prognosis and therapeutics
Introduction
IHC interpretation
Protocol for PDL-1 IHC
Agient Dako kit contains following reagents
Scoring of PDL-1
Chapter Ten. Molecular markers based on targeted therapy in different tumors
Introduction
Breast cancer
Lung cancer
Colorectal cancer
Melanoma
Prostate cancer
Molecular pathways for targeted therapy in cancers
Marker detection method in targeted therapies
Advantages over traditional chemotherapy
Clinical utility and success stories
Future perspective
Personalized treatment approaches
Liquid biopsies for real-time monitoring
Immunotherapy biomarkers
Integration of Artificial Intelligence (AI)
Conclusion
Chapter Eleven. Circulating tumor cell biomarkers and exosomal markers: Advances and laboratory approaches
Introduction
The flow diagram shows the biogenesis of exosomes (Fig. 11.1)
Exosomes as diagnostic and predictive biomarkers for cancer
Nucleic acids in cancer exosomes
Exosomal noncoading RNAs/microRNAs
Exosomal circRNAs
Proteins in cancer exosomes
Hypoxia
Cancer cell–derived exosomes
Detection methods of CTCs and exosomal markers
Future perspectives
Conclusion
Index

Seema Nayak

Dr Seema Nayak has been a Scientist of Molecular Biology and Immunology at Medanta Hospital, Lucknow since 2019. She received the women scientist fellowship from ICMR HRD-DHR in 2015. She also worked as a Research Officer in the Department of Pathology at King George’s Medical University, Lucknow. She has vast experience in oncology research, biomarkers, advanced Genetics, epigenetics, and molecular and biochemical techniques. Her PhD work was focused on biomarker research in oncology. Her post-doctoral work was on Genomics in Cancer Biomarkers. She has contributed high impact publications in PLOsOne, Oral Oncology journals and in cancer biomarkers and oncology. She is a member of several national and international academic organizations and a reviewer in various international peer reviewed journals. She has been actively involved in cancer diagnostics since 2007. She was the pioneer in starting Cancer biomarker diagnostics based on Immunohistochemistry in India.

Affiliations and expertise

Scientist of Molecular Biology and Immunology, Department of Pathology and Laboratory Medicine, Medanta Hospital, Lucknow, Uttar Pradesh, India

Anupam Kumar Sriwastava

Dr Anupam Kumar Sriwastava is a Demonstrator in the department of Biochemistry in the All India Institute of Medical Sciences Bathinda, Punjab since 2020. He has a PhD in Biomedical Sciences from King George’s Medical University, Lucknow. He served as Research coordinator in different Phase III Clinical trials and as university representative in entrance exams conducted by KGMU. During his doctoral he did research in reconnoitre and analysis of diagnostic biomarkers for diagnostic and prognostic purposes in urine of cancer patients and published articles in world reputed journals. He also worked as Assistant Professor in Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur Chhattisgarh and educated the students of B Sc and M Sc classes in different areas of Biotechnology.

Affiliations and expertise

Demonstrator, Department of Biochemistry, All India Institute of Medical Sciences, Bathinda, Punjab, India

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