Question 14: Were the cells in the infused product selected / modified / engineered prior to infusion?
Select Yes if the cells contained in the product were selected (i.e., selective retention of a population of desired cells through recognition of specified characteristics), modified or genetically engineered.
Question 15: Specify the portion manipulated
If the product being infused as a cellular therapy is a portion from a prior HCT (e.g., donor cellular infusion), the portion becomes the “entire” product for the purposes of this form. The product can then be further divided.
Indicate if the Entire product or a Portion of the product was manipulated.
Question 16: Was the unmanipulated portion of the product also infused?
Indicate Yes or No if the unmanipulated portion of the product was also infused.
Question 17: Was the same manipulation method used on the entire product / all portions of the product?
Indicate Yes or No if the same manipulation was used on the entire product or all portions of the product. All manipulations for each portion of the product should be reported in the next questions.
Questions 18-19: Specify method(s) used to manipulate the product (check all that apply)
Indicate the method(s) of manipulation.
Cultured (ex-vivo expansion): Cells were placed in culture to increase in number (i.e., to expand) allowing for sufficient cells for infusion.
Induced cell differentiation: Cells were placed in culture to give rise to cellular elements with biological characteristics other than those of the cells being cultured (i.e., mesenchymal stromal cells cultured to make osteoblasts; pluripotent stem cells cultured to make neural cell precursors). Usually, the description of the process would include the term “differentiation of cells X into cells Y”. This scenario can be seen in regenerative medicine indications.
Cell selection – positive: The manipulation of a cellular therapy product that a specific cell population(s) is enriched. This may be achieved by using an antibody that binds to a specific population of cells (e.g., CD4+ selection).
Cell selection – negative: The manipulation of a cellular therapy product such that a specific cell population(s) is reduced.
Cell selection based on affinity to a specific antigen: The cellular product undergoes selection to isolate the target population based on the ability of the target population to bind or recognize a specific antigen (e.g., a T cell population recognizing viral proteins, or a protein associated with a cancer).
Genetic manipulation (gene transfer / transduction): Cells are manipulated via gene transfer, a process by which copies of a gene are inserted into living cells to induce synthesis of the gene’s product; or transduction, a process by which foreign DNA is introduced into a cell by a virus or viral vector. These techniques deliberately alter the genetic material of an organism to make them capable of making new substances or performing new or different functions. Report the types of genetic manipulation(s) below.
If a method of manipulation was performed on the product, but is not listed above, select Other cell manipulation and specify the manipulation.
Questions 20-28: Specify the type(s) of genetic manipulations (check all that apply)
Select all types of genetic manipulations that apply to the cell therapy product.
Viral transduction is a process by which nucleic acid (DNA) is introduced into a cell by a virus, followed by viral replication in the affected cell.
- Lentivirus: Lentiviruses are members of the genus of retroviruses that have long incubation periods and cause chronic, progressive, usually fatal disease in humans and other animals.
- Retrovirus: Retroviruses are any group of RNA viruses that insert a DNA copy of their genome into the host cell to replicate. HIV is an example of a Retrovirus.
- Transposon: Transposons are discrete mobile sequences in the genome that can transport themselves directly from one part of the genome to another without the use of a vehicle such as phage or plasmid DNA. They move by making DNA copies of their RNA transcripts which are then incorporated into the genome at a new site.
Non-Viral transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells.
- Electroporation: Electroporation is a process of introducing DNA or chromosomes into cells using a pulse of electricity to briefly open the pores in the cell membranes.
- Other non-viral transfection: A different non-viral transfection method not previously listed was utilized. Specify the other non-viral transfection method.
- Gene editing is a type of genetic engineering in which DNA is inserted or removed from a genome using artificially engineered nucleases. If gene editing is selected, select all applicable genes edited in the manipulation. If an Other gene was edited, specify the other gene.
Non-native protein expression is a type of genetic engineering in which a gene is transferred codes for an antigen receptor other than one that may already be naturally present in the cell (e.g., T-cells have natural T-cell receptors [TCRs]; a transgenic TCR or a Chimeric Antigen Receptor [CAR] are non- native antigen receptors).
- Chimeric Antigen Receptor (CAR): A cell-surface receptor that has been engineered to combine novel features and specificities from various sources to enhance its antigen specificity. Engineered T- cells or B-cells will produce the specialized receptor that will be capable of binding to an epitope on its target cell1.
- The CAR construct consists of several genes that can exert different functions, such as augment the immune response by co-stimulation, increase affinity, and increase the time it persists in the circulation without being cleared. The CAR construct information is usually unique and may influence its effect against the disease or the severity of side effects. Specify which construct(s) was used in the making of the Chimeric Antigen Receptor (CAR). If a construct was utilized that is not in the list, check Other construct and specify the other construct.
- CD19bζ (zeta) is an antibody fused to CD3ζ (zeta) and should be reported as CD3ζ.
For more information related to the different constructs and their functions, see this article:
https://www.jci.org/articles/view/80010.
1NCIthesaurus: https://ncit.nci.nih.gov/ncitbrowser/
Suicide gene: Cells underwent manipulation to have cell suicide inducing transgenes inserted into the product. Select all suicide genes that apply to the cell therapy product. If Other suicide gene is selected, specify the other suicide gene.
iCasp9 is inducible Caspase 9. CaspaCIDe® consists of an inducible caspase 9 (iCasp9) gene together with the small-molecule, bio-inert, chemical induction of dimerization (CID) drug, AP1903.
T-cell receptor: genetically modifying a recipient’s T cells to express T cell Receptors (TCRs) that target a specific cancer cells.
If another type of genetic manipulation was performed that is not listed above, select Other genetic manipulation and specify the other genetic manipulation.
Question 29: Was the product manipulated to recognize a specific target/antigen?
Indicate Yes if the cells were cultured or engineered so that the majority of cells in the end product are able to recognize or bind to a chosen target (e.g., proteins from a virus or a protein from a tumor). This manipulation can be done outside of the context of a ‘genetic manipulation’.
Questions 30 – 35: Specify target (check all that apply)
Select all targets appliable to the cell therapy product: Viral, Tumor / cancer antigen, or Other target.
If the target is Viral, select all target viral antigen(s) that apply to the product.
If the target is Tumor / cancer antigen, select all target tumor / cancer antigen(s) that apply to the product.
If the target is something other than viral or tumor/cancer antigen, select Other target and specify the other target.
Section Updates:
| Question Number | Date of Change | Add/Remove/Modify | Description | Reasoning (If applicable) |
|---|---|---|---|---|
| . | . | . | . | . |
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