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EP-4737550-A1 - CELL ENGINEERING TARGET CELLS IN A MIXTURE COMPRISING THE TARGET CELLS AND NON-TARGET CELLS

EP4737550A1EP 4737550 A1EP4737550 A1EP 4737550A1EP-4737550-A1

Abstract

In a first aspect, the present invention relates to a system (1) for cell engineering target cells (51) in a mixture (5) comprising the target cells and non-target cells (52), comprising: i) a cell monitoring component (41) for tracking and classifying individual cells (50) in the mixture (5), wherein classifying a cell (50) comprises determining whether the cell (50) is a target cell (51); ii) an engineering component (42) for selectively modifying individual cells (50) at one or more engineering sites (420), wherein the target cells (51) and the non-target cells (52) are randomly distributed in the engineering component (42); and iii) a control component (43) for controlling the engineering component (42) so as to selectively modify only target cells (51) tracked to be at one of the engineering sites (420).

Inventors

  • STAKENBORG, TIM
  • Rocco, STIRPARO
  • Peter, Peumans

Assignees

  • Imec VZW

Dates

Publication Date
20260506
Application Date
20241031

Claims (15)

  1. A system (1) for cell engineering target cells (51) in a mixture (5) comprising the target cells (51) and non-target cells (52), comprising: i) a cell monitoring component (41) for tracking and classifying individual cells (50) in the mixture (5), wherein classifying a cell (50) comprises determining whether the cell (50) is a target cell (51); ii) an engineering component (42) for selectively modifying individual cells (50) at one or more engineering sites (420), wherein the target cells (51) and the non-target cells (52) are randomly distributed in the engineering component (42); and iii) a control component (43) for controlling the engineering component (42) so as to selectively modify only target cells (51) tracked to be at one of the engineering sites (420).
  2. The system (1) according to any of the previous claims, wherein selectively modifying individual cells (50) comprises selectively inserting and/or modifying a nucleic acid in the individual cells (50).
  3. The system (1) according to claim 2, wherein selectively modifying the individual cells (50) comprises selectively transfecting the individual cells (50).
  4. The system (1) according to claim 3, wherein transfecting comprises electroporating, photoporating, cell squeezing, magnetofecting, chemical transfecting or biochemical transfecting.
  5. The system (1) according to any of the previous claims, wherein a composition of the mixture (5) is maintained between classifying the individual cells and selectively modifying the individual cells.
  6. The system (1) according to any of the previous claims, wherein the cell monitoring component (41) comprises an imaging unit (415), preferably a computational microscopy unit, most preferably a lens-free imaging unit.
  7. The system (1) according to any of the previous claims, wherein the cell monitoring component (41) comprises a fluidic channel (6) comprising a cell movement modifying surface (411).
  8. The system (1) according to claim 7, wherein the fluidic channel (6) comprises one or more structures having the cell movement modifying surface (411).
  9. The system (1) according to claim 7 or 8, wherein the fluidic channel (6) comprises multiple subzones (413, 414) having distinct cell movement modifying surfaces.
  10. The system (1) according to any of the previous claims, wherein determining whether the cell (50) is a target cell (51) is based on a determination of the shape, size, transparency, scattering, speed, direction and/or path of the cell (50).
  11. The system (1) according to any of the previous claims, wherein the target cells (51) are stem cells or immune cells, preferably peripheral blood mononuclear cells, more preferably lymphocytes, most preferably T cells or NK cells.
  12. A method for cell engineering target cells (51) in a mixture (5) comprising the target cells (51) and non-target cells (52), comprising: a) classifying individual target cells (51) in the mixture (5), wherein classifying a cell comprises determining whether the cell (50) is a target cell (51); b) tracking at least some of the target cells (51); and c) selectively modifying at least some of the tracked target cells (51), wherein the target cells (51) and the non-target cells (52) are randomly distributed.
  13. The method according to any of claim 12, wherein step b-and optionally step a-comprises computational imaging of the individual target cells (51).
  14. The method according to claim 12 or 13, wherein selectively modifying at least some of the tracked target cells (51) comprises selectively inserting and/or modifying a nucleic acid in said the tracked target cells (51).
  15. The method according to any of claims 12 to 14, wherein a composition of the mixture (5) is maintained throughout step a, b and up to c.

Description

Technical field of the invention The present invention relates to the field of cell engineering, and more specifically to selectively modifying individual target cells within a mixture of target and non-target cells. Background of the invention In the field of cellular biology and medical research, the ability to modify (e.g. genetically) cells at the single-cell level is crucial e.g. for advancements in therapy and for furthering our understanding of cellular functions. Traditional approaches to modifying target cells in a mixture of cells typically involve sorting out the target cells, followed by transduction or transfection of the target cells. These approaches thus typically require the physical separation of target cells from a heterogeneous mixture, which can be inefficient and yield less than optimal results. Moreover, these approaches generally handle cells in batches rather than on an individual basis, limiting control in cell modification tasks. Additionally, sorting is generally done using label-based methods, such fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS). These techniques, while effective for certain applications, require the cells to be labelled (e.g. with fluorescent markers for FACS or magnetic beads for MACS). This labelling process not only adds additional steps (e.g. additional sample preparation and wash steps to remove unbound labels or reagents) to the procedure-making it time-consuming, complex and more at risk of cell loss or damage-but also introduces foreign materials that can alter cell behaviour/function and/or cell viability. These issues become exceedingly critical when dealing with rare cell types or when sample sizes are limited. One area in which cell engineering is particularly useful is that of cell therapy. Our immune system plays a pivotal role in defending the body against infections and diseases. It possesses the remarkable ability to adapt and respond to various pathogens and internal threats, including viruses, bacteria, cancerous cells, fibrosis and aging. However, there are instances where the immune system fails to make the right tool to fight back (e.g. for HIV), is too slow and needs to be prepared upfront (e.g. with vaccines), or struggles when the threat is an alteration of our own cells (e.g. in the case of cancer or aging). To bolster the immune system's capacity to fight diseases, various strategies have been developed to prime the immune system for fighting a disease of interest. For example, vaccination is a well-known method that prepares the immune system in advance to combat infections, such as influenza or COVID-19. Another advanced approach involves cell therapies, such as chimeric antigen receptor T (CAR-T) cell therapies, which entail modifying the receptors of T-cells to specifically bind to a particular target antigen on certain cells (e.g. cancer cells). While CAR-T therapies have demonstrated promising results, especially in treating previously untreatable cancers, the production of these therapies is fraught with challenges. The current manufacturing processes for cell therapies are complex and time-consuming, often requiring several weeks to produce a single dose. This involves multiple steps such as cell extraction, selection, modification, and expansion, each necessitating specialized equipment, facilities and personnel. Furthermore, extensive quality control tests at each stage contribute to the overall cost and duration of therapy production. The logistics involved further complicate accessibility, as most manufacturing facilities are centralized, necessitating the transportation of biological materials over long distances, potentially compromising their viability and efficacy. Additionally, cell engineering is also extensively used in lab/research settings; e.g. in gene expression studies, knock-down/out studies, protein production, stem cell research, high-throughput screening, miRNA studies, etc There is thus still a need in the art for ways to modify target cells which address at least some of the issues outlined above. Summary of the invention It is an object of embodiments of the present invention to provide an apparatus for selectively modifying individual target cells in a mixture comprising the target cells and non-target cells. This objective is accomplished by systems, modules, cartridges and methods according to the present invention. The claimed invention is as set out in the appended claims. It is an advantage of embodiments of the present invention that they are capable of selectively modifying target cells, ensuring that only the targeted cells are modified. It is a further advantage of embodiments of the present invention that they allow for the modification of target cells without the necessity of isolating them from a mixture comprising both target and non-target cells. It is yet a further advantage of embodiments of the present invention that they can enable the classificatio