Cellular Radiation Effects

Cell membrane - Alteration in permeability Cellular organelles - Functional Aberrations Nuclear membrane - Altered permeability & Function DNA - Chromosomes - Functional aberrations

DNA (Chromosomes)
The DNA makes up the chromosomes of the cell and carries all of the functional encoding information of the cell or organism All of the chromosomes together make up the genome The genome is composed of many genes (60,000 in humans) The individual genes are composed of sequences of nitrogenous bases attached to the molecular backbone. These sequences encode for protein functions etc. which control all cell functions Large areas of a DNA strand may not be expressed in individual cells

DNA Structure
Double stranded helix (twisted ladder millions of rungs long) with side rails of ladder composed of Sugar molecules bound together by a phosphate Rungs are composed of the nitrogenous bases Adenine, Thymine, Guanine and Cytosine. Adenine and Thymine combine to make up one type of rung and Guanine and Cytosine combine to make up another type. A given base may be on either side of the helix

DNA Structure
DNA is a very large molecule. There are about 2 x 109 base pairs in the mammalian genome distributed across 15-100 chromosomes.

The stearic configuration (shape) of the molecule changes constantly and is important to function.
DNA is replicated at cell division

DNA Structure

DNA Structure

Mechanism of radiation Injury

Direct ionization of a portion of the DNA molecule. Indirect injury by free radicals in the DNA environment.
H+, 0H-, H202-, etc.

Mechanism of radiation Injury

DNA Radiation Injuries

Base pair deletion Cross-linking injuies Single Strand Break Double Strand Break Multiple (complex) lesions

DNA Radiation Injuries

DNA Radiation Injuries

DNA Replication
DNA is replicated during S Phase prior to the onset of mitosis The original DNA is used as a template for the building of the new DNA.

Quite rapid process, requires less than 15 hours.

DNA Replication

Cell Division
Mitosis
Multistep process
DNA organizes into identifiable chromosomes (Prophase ) DNA aligns with centromeres on equatorial plate (Metaphase) DNA Separates and moves to opposite ends of cell (Anaphase) Cell cytoplasm divides at equatorial plate (Telophase)

Cell Division

Mitosis
Cell resumes normal functional operations (interphase)

Through this process radiation induced aberrations in the DNA may result in significant loss of DNA to one or both of the daughter cells.
Only requires about one hour

Radiation Induced Chromosomal Aberations

Chromatid exchanges. Sister Unions Acentric Fragments Rings Dicentric Unions

Radiation Induced Chromosomal Aberations

Radiation Induced Chromosomal Aberations

Cell Cycle
Tissues grow and are maintained through cell replication (regeneration) Some cells never divide once adulthood is reached. There are a specific set of steps involved
G1 (G0) S G2 M Gap Phase 1 Synthesis Gap phase 2 Mitosis Functional cell DNA synthesis Rest Cell Division

Cell Cycle

Repair of Radiation Injury

Cellular mechanisms are in place which can repair most if not all types of radiation injury to the DNA. Repair is a time sensitive process Repair is a cell cycle dependent process Repair is a dose rate dependent process Repair is dose dependent Repair is radiation type dependent

Cellular Mechanisms of Repair

Base Excision Repair
Damaged bases must be repaired The complementary base on the opposite strand serves as a template. This type of repair is quite efficient Loss of this repair mechanism increases the incidence of mutations.

Cellular Mechanisms of Repair

Nucleotide Excision Repair (NER) Repairs DNA damage due to pyrimidine dimer adducts added to the DNA by injury. - Enzymatic removal of lesion and associated backbone. - Lesion is then sealed by DNA polyemerase and ligase. - Defective mechanism increases sensitivity to UV light

Cellular Mechanisms of Repair

Double Strand Break Repair
Nonhomologous End Joining
Occurs primarily in S phase when no sister chromatid is present. In some instances the base pair sequence is filled in by repair processes without a template. Complex process with multiple pathways Because it is an error prone process it tends to promote development of mutations.

Cellular Mechanisms of Repair

Double Strand Break repair
Homologus Recombination repair
Uses sister chromatid as a template to faithfully recreate the damage section and join the ends together properly Occurs in G1 phase when sister chromatids present Error free process Loss of ability increase radiation sensitivity and mutation rate.

Cellular Mechanisms of Repair

Single strand break repair Occurs via similar pathway to Base Excision Repair. Efficiently done and vast majority of lesions are repaired.

Cellular Mechanisms of Repair

Because of the efficiency of repair mechanisms for all but double strand breaks the majority of the cell killing occurring at low doses is due to double strand breaks which are not repaired. At high doses accumulated DNA injury due to many single strand breaks and base pair deletions becomes more important.

Types of DNA Damage

Lethal Damage
Irreversible and irreparable fatal to cell

Potentially Lethal Damage (PLD

Damage which is lethal unless modified by post irradiation events

Sublethal Damage (SLD)

Repairable injury to the DNA

Lethal Damage
Non repairable injury associated with double strand breaks Increases with LET up to a point Increases with higher doses

Potentially Lethal Damage

Not repaired and lethal under normal circumstances. Repair increased by conditions which are suboptimal to the division of the cell
Reduced temperature Hypoxia Low pH Others

Increased capability = radioresistance

Sublethal Damage Repair (SLD)

Refers to DNA damage that is repaired Splitting radiation dose increases survival Occurs in 1-6 hours after irradiation Affected by phase of cell cycle Affected by cell cycle time
Long cycle usually increases repair

Indicated by shoulder on survival curve

Repair is a time sensitive process

Repair of DNA injury of all types is essentially complete by 6 hours post irradiation. External factors that affect cellular metabolic rate may delay or accelerate it Foundation of modern radiotherapy

Repair is a cell cycle dependent process

Different phases have different repair capabilities
Mitosis has the least repair capability G2 G1/G0 S phase has the most repair capability

Capability varies in G1 and S

S-phase Radiation Resistance

Likely due to Homologous Recombination Can result in cell population synchrony
S G2 blockade and increased survival in S

More important in rapidly dividing cells May be important in some tumor lines

Reassortment
Cells in G2 & M are preferentially killed Cells in S are preferentially spared. Alters proportion of cells in each phase Cell population tends to reestablish normal proportions within 2-3 cycles. Killed cells replaced by cells from G1 Moves cells to more sensitive G2 & S

Repair - dose rate dependency

Dose rate decreased by two mechanisms
Splitting dose into smaller fractions w/ time between the fractions
Smaller fractions increase time if spacing constant

Reducing the actually rate at which dose is delivered

Repair between ongoing during doses Repopulation may occur

Repair is dose rate dependent

At very low dose rates repair of SLD can keep up with radiation damage.
SLD predominate type of injury.

Repopulation can account for LD and SLD

Dependent on cycling cell population Cell cycle time short relative to dose rate

Affected by radiation quality Mutation rates may be increased

Repair - dose rate dependency

Repair - dose rate dependency

Repair is dose dependent

Lethal Damage increase with dose PLD increases with dose Accumulation of SLD increase with dose Survival curve is continuously bending
Some repair always present Various forms of damage interact

Repair is radiation type dependent

Low LET radiation is repaired Little repair of High LET radiation injury
Dense ionization track Double strand breaks more likely Energy deposition curve dependent

Sublethal damage less important

Single strand breaks, base pair deletion, etc.

LET vrs. Survival