Oncogenes and Stem Cells

Oncogenes

  • We're finishing our discussion of oncogenes and moving into stem cells.
  • The presence of an oncogene, such as Sark, causes cancer.
  • This was discovered by Temin in 1910 with Rous sarcoma virus.
  • Later studies in the 1970s-1980s used recombinant DNA to create Delta RSV.
  • Sark was present in all chicken cells, so the mutation of C-Sarc (a normal tyrosine kinase regulating cell growth) into V-Sarc (a constitutive kinase with constitutive signaling) causes cancer.
  • Michael Bishop and Harold Barmus won the Nobel Prize for discovering this.
  • Examples of proto-oncogenes being converted into oncogenes -- genes that you turn on, but don't turn off, like Ras.
  • Another type of mutation that causes cancer is the loss of a tumor suppressor gene like p53, which prevents damaged DNA from being replicated.
  • Losing p53 functionality is bad for both initial cancer development and making it worse.
  • Tumors acquire multiple mutations by the time cancer kills you.
  • Half of cancer patients have lost p53.
  • We have 2 copies of every gene so we don't have a problem until we lose both functional copies of p53.
  • Tumor suppressor genes are recessive, while oncogenes are dominant.

Mutations that Cause Cancer

  • Types of mutations that generate cancer:
    • Coding mutation: changes the sequence of the gene.
      • Introduce a mutation in the DNA.
      • The RNA sequence is incorrect.
      • The protein is hyperactive (e.g., Ras never turns off).
    • Gene amplification: more copies of the DNA.
      • Nothing wrong with the gene sequence.
      • More copies of DNA result in more RNA and more protein.
    • Chromosome rearrangement: a new promoter is placed in front of the DNA.
      • The gene itself is not mutated, nor is its quantity.
      • The new promoter is more active, resulting in more RNA and more protein.
    • Fusion protein: recombination in the DNA causes chunks of DNA to move around, resulting in two proteins glued together.
      • RNA comprises two proteins glued together.
      • A new piece gets glued on yielding a hyperactive protein.
  • Too much growth signal could be sent by proteins in some cases with other cases involving merely too much protein.

Overexpression of the Egf Receptor

  • Instead of a normal density of Egf receptors, you get many.
  • You can have 10 to 50 times the normal levels of Egf receptor.
  • The receptors are super crowded, leading to autophosphorylation even without the Egf ligand present.
  • This cross-phosphorylation turns on the cell and tells it to grow, even without Egf, leading to growth factor independence
  • Autocrine signaling is another growth factor independence mechanism.
  • A cell gains the ability to secrete its own mitogen (Egf), telling itself to grow regardless of neighbors' signals.

Truncated Egf Receptor

  • A truncated Egf receptor has a mutation without the part that binds the Egf ligand.
  • The truncated Egf receptor turns on regardless of whether Egf is there and this receptor cannot even bind to Egf.
  • Erisa is a lung cancer drug that targets a specific type of truncated Egf receptor, turning it off.
  • Erisa works great in 10% of patients, there is a dramatic reduction in the tumor, but does nothing for the other 90%.
  • We can analyze the genetics of a tumor to see if the patient has this mutation and would benefit from Erisa, an example of patient-specific medication.

Stem Cells

  • Stem cells were first understood in the 1960s with the discovery of hematopoietic stem cells.
  • Radiation effects were studied after nuclear bombs were created, when scientists realized they didn't know the effects of radiation exposure.
  • Mice were irradiated and died in around 30 days. Bone marrow transplants were discovered and rescued the irradiated mice so that they didn't die.
  • As few as 10,000 bone marrow cells were enough to rescue a mouse.
  • Sometimes it worked with fewer than 10,000 cells, but sometimes those transplants failed, it was very binary.
  • Just one cell was enough to rescue the mouse.
  • Clones of this single cell could also rescue other mice.
  • This magic cell is the hematopoietic stem cell, which can turn into long-term and short-term hematopoietic stem cells.
  • The short-term hematopoietic stem cells will create the myeloid lineage (red blood cells, platelets, granulocytes, macrophages) and the lymphoid lineage (B cells, T cells, natural killer cells) and the dendritic cells -- every single blood cell type.
  • The single transplanted cell can rescue the entire blood supply.
  • The long-term hematopoietic stem cell can differentiate into the short-term, and then the myeloid and lymphoid lines of cells, and it can also clone itself via self-renewal.
  • The short-term hematopoietic stem cells can also self-renew, but do it faster.
  • Hematopoietic stem cells are needed to make new blood; without them, you can't make new blood.
  • Blood cells, especially red blood cells, last about 120 days before they are replaced.

Stem Cell Balance

  • Stem cells make new differentiated cells, which are the myeloid and lymphoid cells.
  • Stem cells must also self-renew.
  • The stem cells need to maintain a 1:1 balance between self-renewal and differentiation.
  • Too much self-renewal leads to a buildup of stem cells, if there too little self renewal, we risk losing the population.
  • The hematopoietic stem cells have slow cycling (copy DNA less often), while short-term hematopoietic stem cells undergo fast cycling in the rapidly dividing state.
  • Fast cycling means racing through the cell cycle as fast as possible.
  • A typical human cell completes a cell cycle in about 24 hours.
  • Every time DNA is copied, there is a chance of error.
  • Even with error-checking machinery, one to three errors can be missed each duplication.
  • Errors can lead to uncontrolled growth and cancer.
  • Stem cells use slow cycling to maintain high-quality DNA.
  • Short term stem cells are produced to take care of any short fall of not replenishing the blood supply sufficiently.

Other Stem Cell Types

  • Stem cells exist in many tissues, aiding in healing when there is damage.
  • Other stem cell types include:
    • Intestines
    • Skin and hair (generated by the same stem cell)
    • Bone and cartilage (mesenchymal stem cell)
    • Neural stem cells (limited ability to make new neurons)
    • Cardiac stem cells (limited ability to renew heart tissue)
  • Hematopoietic stem cells and mesenchymal stem cells are multipotent, making a bunch of different cell types.
  • Pluripotent stem cells can make anything.
  • The intestinal lining takes a lot of damage due to consumption of various foods and exposure to carcinogens, and requires renewal every 3 to 4 days.

Intestinal Stem Cells

  • In the intestines, intestinal crypts exist with a handful of stem cells at the bottom, away from the surface.
  • Next to the stem cells are other cells that help the stem cells stay as themselves.
  • Further up the crypt, the stem cells turn into transit amplifying cells that cycle quickly.
  • The stem cells cycle slowly, while the transit amplifiers cycle quickly.
  • The villi are finger-like protrusions that stick up into the intestines to absorb nutrients.
  • Mucus-secreting cells create a lining of mucus.
  • Absorptive cells absorb nutrients.
  • Both mucus-secreting cells and nutrient absorbers come from the stem cell in absorptive cells.
  • The cells start off on the bottom, move up, become transit amplifiers, and then differentiate into fully differentiated cells.
  • The cells walk up the villi, and when they get to the end, they undergo programmed cell death.
  • Colon cancer has a chance to develop, but these defense mechanisms are in place to prevent it from happening.

Homeostasis

  • Maintaining proper cell number requires homeostasis.
  • Feedback control mechanisms are important for cell number regulation, like maintaining blood volume or intestine size.

Blood Regulation

  • The body has about 10 liters of blood, with 5 \times 10^{12} red blood cells per liter.
  • The process of making 10^{10} red blood cells every hour maintains this system.
  • At sea level, the amount of oxygen is normal; at higher altitudes the concentration declines.
  • If people spend a few days in a location with low oxygen concentration such as in the mountains in Mammoth, California, their body adjusts.
  • Denver, Colorado is known as the 'mile high city'. The people from Denver practice and live in low oxygen concentrations, thus conditioning them when competing with athletes in sea level conditions.
  • Erythropoietin (Epo) is an endocrine signal secreted into the blood for red blood cell control.
  • If blood oxygen is low, the kidneys release Epo.
  • Epo slows down the rate at which red blood cells decompose, increasing the blood oxygen level, giving the body time to adjust and acclimate to the higher altitudes.
  • Injecting recombinant Epo is a form of illegal doping, particularly among endurance athletes, thus, Epo is banned in many events.