Emerging Biophotonic Technologies in the Diagnosis and

Emerging Biophotonic Technologies in the Diagnosis and Treatment of Diseases

Philip Chen, MD, PhDCognoscenti Health Institute

UCF/CREOL4/1/2005

What is Biophotonics?

νν BiophotonicsBiophotonics is the science of generating and is the science of generating and harnessing light (photons) to image, detect and harnessing light (photons) to image, detect and manipulate biological materials.manipulate biological materials.

νν BiophotonicsBiophotonics is used in is used in BIOLOGYBIOLOGY to probe for to probe for molecular mechanisms, function and structure. molecular mechanisms, function and structure. It is used in It is used in MEDICINEMEDICINE to study tissue and to study tissue and blood at the macro (largeblood at the macro (large--scale) and micro scale) and micro (very small scale) organism level to detect, (very small scale) organism level to detect, diagnose and treat diseases in a way that are diagnose and treat diseases in a way that are nonnon--invasive to the body. invasive to the body.

NSF Center for NSF Center for BiophotonicsBiophotonics, Science and Technology at UCSD, Science and Technology at UCSD

1674

Anton Van Leeuwenhoek observed bacteria, yeast and other living microorganisms under the microscope.

1665

Robert Hooke observed cells under the microscope.

Charles A. SpencerNew York, 19th Century

“With Spencer's glasses I was able, without difficulty, to see the cross lines on the Naviculahippocampus, while with Chevalier's, by the same light, I could see them only with great difficulty.”J.W. Bailey

Prof. of Chemistry, U.S. Military AcademyThe American Journal of Science, Nov 27, 1841

1931

Ruska and Knott invented Electron microscope. They were awarded Nobel Prize in 1986 for this invention

1965

First production Scanning Electron Microscope, the Stereoscan was introduced by Charles Oatley

““LightsLights”” in Medicinein Medicineνν Radiology Radiology –– XX--ray, MRI, CT Imagingray, MRI, CT Imagingνν LASIK SurgeryLASIK Surgeryνν Laser UrologyLaser Urologyνν Laser Plastic SurgeryLaser Plastic Surgeryνν Laser tissue solderingLaser tissue soldering--

””biological gluebiological glue””νν EndoscopicEndoscopic Laser OtolaryngologyLaser Otolaryngologyνν Laser DentistryLaser Dentistryνν Radiotherapy Radiotherapy –– Photons and Photons and

UltrasoundUltrasoundνν Clinical Laboratory Spectroscopy and Clinical Laboratory Spectroscopy and

MicroscopyMicroscopyνν Genomic Medicine Genomic Medicine –– Diagnosis and Diagnosis and

TreatmentTreatment

Genomic Medicine

ν Diagnoseν Predict Disease

Progressionν Treatν Monitor treatment

safety and efficacy

DNA-based Testing

ν Presymptomatic testing for predicting adult-onset disorders (e.g., Huntington's disease)

ν Presymptomatic testing for estimating the risk of developing adult-onset cancers and Alzheimer's disease

ν Confirmational diagnosis of a symptomatic individual (e.g., Factor V Leiden)

DNA based testing

ν Prenatal diagnostic testing (e.g., Cystic Fibrosis)

ν Newborn screening

ν Embryos Screening (pre-implantation)

DNA based testing

ν Carrier screening - identify unaffected individuals who carry one copy of a gene

ν Forensic/identity testing (i.e., paternity test)

Alpha-1-antitrypsin deficiency (AAT; emphysema and liver disease) Amyotrophic lateral sclerosis (ALS; Lou Gehrig's Disease; progressive motor function loss leading to paralysis and death)

Alzheimer's disease* (APOE; late-onset variety of senile dementia) Ataxia telangiectasia (AT; progressive brain disorder resulting in loss of muscle control and cancers)

Gaucher disease (GD; enlarged liver and spleen, bone degeneration) Inherited breast and ovarian cancer* (BRCA 1 and 2; early-onset tumors of breasts and ovaries)

Hereditary nonpolyposis colon cancer* (CA; early-onset tumors of colon and sometimes other organs) Charcot-Marie-Tooth (CMT; loss of feeling in ends of limbs)

Congenital adrenal hyperplasia (CAH; hormone deficiency; ambiguous genitalia and male pseudohermaphroditism) Cystic fibrosis (CF; disease of lung and pancreas resulting in thick mucous accumulations and chronic infections)

Duchenne muscular dystrophy/Becker muscular dystrophy (DMD; severe to mild muscle wasting, deterioration, weakness) Dystonia (DYT; muscle rigidity, repetitive twisting movements)

Fanconi anemia, group C (FA; anemia, leukemia, skeletal deformities) Factor V-Leiden (FVL; blood-clotting disorder)

Fragile X syndrome (FRAX; leading cause of inherited mental retardation) Hemophilia A and B (HEMA and HEMB; bleeding disorders)

Hereditary Hemochromatosis (HFE; excess iron storage disorder) Huntington's disease (HD; usually midlife onset; progressive, lethal, degenerative neurological disease)

Myotonic dystrophy (MD; progressive muscle weakness; most common form of adult muscular dystrophy) Neurofibromatosis type 1 (NF1; multiple benign nervous system tumors that can be disfiguring; cancers)

Phenylketonuria (PKU; progressive mental retardation due to missing enzyme; correctable by diet) Adult Polycystic Kidney Disease (APKD; kidney failure and liver disease)

Prader Willi/Angelman syndromes (PW/A; decreased motor skills, cognitive impairment, early death) Sickle cell disease (SS; blood cell disorder; chronic pain and infections)

Spinocerebellar ataxia, type 1 (SCA1; involuntary muscle movements, reflex disorders, explosive speech) Spinal muscular atrophy (SMA; severe, usually lethal progressive muscle-wasting disorder in children)

Thalassemias (THAL; anemias - reduced red blood cell levels) Tay-Sachs Disease (TS; fatal neurological disease of early childhood; seizures, paralysis) [3/99]

Pharmacogenomics(personalized medicine)

Pharmacogenomics(personalized medicine)

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Pharmacogenomics

ν Viral genomics ν Bacterial genomics

Genotyping : nested RTGenotyping : nested RT--PCRPCR

RT HeatHeat

plasma HIV

Cellular HIV

HeatHeat

Genotyping : Genotyping : ddNTddNT sequencingsequencing

Genotyping : Genotyping : GeneChipGeneChip technologytechnology

K103N mutation in HIV Reverse Transcriptase - confers resistanceto all non-nucleoside reverse transcriptase inhibitors (NNRTIs)

Rationalized drug Rationalized drug and vaccine designand vaccine design

Optical Probes Optical Probes --Single Molecule ImagingSingle Molecule Imaging

Examples: In vitro Examples: In vitro Cellular Imaging to Cellular Imaging to

Target Drug TherapyTarget Drug Therapy

M3 AMLM3 AMLHer2Neu+ Breast CancerHer2Neu+ Breast Cancer

M3 Acute M3 Acute MyelogenousMyelogenousLeukemia (AML)Leukemia (AML)

Submit sample forFlow Cytometry(look at many markerson the cell surface)

Auer Rods! M2, M3, or M4 AML?

If M3 AML

Obtain Chromosome

Analysis (Cytogenetics)

If t(15;17) - Treat with all-trans retinoic acid (ATRA)Retinoic acid binding protein -> oncogene

Flow Flow CytometryCytometry

her2neuher2neu

νν OncogeneOncogene overexpressedoverexpressed in some in some breast cancers.breast cancers.

νν Can stain for overCan stain for over--expression of this expression of this protein on sections of tumor, or look protein on sections of tumor, or look for genetic alterations.for genetic alterations.

νν If positive, treat with If positive, treat with herceptinherceptin -- a a drug that targets the tumor cells.drug that targets the tumor cells.

Flow Flow CytometryCytometry

In Vivo Molecular Imaging

Fluorescent MicroscopyFluorescent Microscopy

ConfocalConfocal MicroscopyMicroscopy

EM/SEMEM/SEM