Preprint
Review

Non-invasive Multiregional Tumor Sampling Using Magnetic Nanoparticle-Loaded Macrophages

Altmetrics

Downloads

963

Views

506

Comments

0

This version is not peer-reviewed

Submitted:

21 August 2023

Posted:

22 August 2023

Read the latest preprint version here

Alerts
Abstract
Some tumors occur in anatomical regions that are hard to biopsy with a needle. Such regions include the brain, spinal cord, liver, and lungs. For the latter two, magnetic nanoparticle-loaded macrophages could be intravenously infused and driven via an MRI machine into the tumor or tumors. Once there, they can be induced to phagocytose whole tumor cells. They would keep their target in a non-digested form by inhibiting phagosome maturation - and be directed via magnetotaxis or chemotaxis to an extraction point in the body where they can be more easily collected via needle.
Keywords: 
Subject: Medicine and Pharmacology  -   Oncology and Oncogenics

1. Introduction

Cancer has plagued multi-cellular organisms since their conception. Recently, I wrote a paper about how many cancer patients may have one or more mutations that are ubiquitous throughout their tumor(s) [1,2,3]. The rest may at least have a small set of subclonal mutations that together cover all sequenced regions of their tumors. These mutations could be targeted by an oncolytic vector with the broadest tropism possible that only replicates and becomes hyper-virulent after detecting said mutations. I called this strategy, “Oncolytic Vector Efficient Replication Contingent on Omnipresent Mutation Engagement” (OVERCOME).
To identify these mutations, multiregion, multi-sample sequencing should be employed for each patient. However, tumors in certain anatomical regions are not easy to biopsy - especially in a multiregional fashion. Such regions include the brain, the spinal cord, the liver, and the lungs.
Here, I explore another way of acquiring multiregion biopsies from tumors that are hard to reach via traditional means.

2. Mechanisms

Bioengineered macrophages could be used for this purpose. They could be loaded with magnetic nanoparticles (MNPs) and steered into the heart or lung tumors using an MRI machine [4]. Perhaps the macrophages should be induced via small molecule to chemorepel [5] each other once they have reached the target site or sites - in order to spread out more evenly throughout the tumor(s). A gene circuit possibly involving ARHGEF37 [6] or Cdc42 could be employed as well - to allow for vigorous, random movement within the tumour in addition to their chemorepulsion from each other.
In either case, once there, they could be induced via small molecule to express a chimeric antigen receptor for phagocytosis (CAR-P) [7]. Alternatively, they can be heated via an alternating magnetic field to induce gene expression of the CAR-P [8]. The CAR-P would target a ubiquitously expressed cell surface protein [9,10]. If necessary, SNIPRs [11] could be utilized to allow for targeting multiple ubiquitously expressed cell surface proteins to ensure phagocytosis of cancer cells throughout the tumor(s).
Inhibiting maturation of and lysosomal fusion with the specific phagosome carrying the target cell could be achieved in a variety of ways [12]. After giving the macrophages some time to collect a target - they would be drawn via magnetism or chemotaxis to an extraction point in the body. Perhaps they can be magnetically drawn to the peritoneal cavity [13] and withdrawn via needle.
Importantly, it was shown that whole cancer cells can be engulfed via this CAR-P method, as opposed to trogocytosis. However, trogocytosis was still more common. Thus, more work may be necessary before this strategy can be employed.
Figure 1 CAR-Ps.
Figure 1. A) The bioengineered macrophages are magnetically steered into the tumor. B) The chimeric antigen receptor is induced, and macrophages phagocytose cancer cells in a patient’s tumor. C) The cancer cell resides within a non-acidified phagosome that does not fuse with a lysosome. D) The macrophages are magnetically drawn to the peritoneal cavity; they are withdrawn via needle. E) The cancer cell genomes are sequenced to look for ubiquitous mutations.
Figure 1. A) The bioengineered macrophages are magnetically steered into the tumor. B) The chimeric antigen receptor is induced, and macrophages phagocytose cancer cells in a patient’s tumor. C) The cancer cell resides within a non-acidified phagosome that does not fuse with a lysosome. D) The macrophages are magnetically drawn to the peritoneal cavity; they are withdrawn via needle. E) The cancer cell genomes are sequenced to look for ubiquitous mutations.
Preprints 82887 g001aPreprints 82887 g001b
An alternative solution would be to install the T-cell lytic granule system in the CAR-P macrophages [14,15]. They would lack granzyme B, but be able to rapidly and directionally secrete perforin to lyse the target cell while sparing the nucleus [16,17]. Then, they could phagocytose debris until they bind the nuclear envelope - and selectively phagocytose that and inhibit the maturation of the phagosome and phagosome-lysosome fusion. (The nucleus would be a smaller target than the cell as a whole.)
Yet another strategy would be for carrier, Irf8(-/-) [18] macrophages to ferry intracellular, phagocytic [19], magnetosome [20]-bearing bacteria to cancer cells. The bacteria would lyse the carrier macrophages once in the tumor region, invade tumor cells, and phagocytose their nuclei. Then, they would lyse the cancer cell - and could be directed chemotactically or magnetotactically to an extraction point.
The macrophages could also simply attach to the cancer cells and magnetotactically pull them to an extraction point. It would be very important that they do not drop their cargo along the way. However, the cancer cell could theoretically replicate while being towed.
Finally, cell-cell fusion [21] could potentially be locally induced in the tumor - as long as the macrophage had many chromosomally-encoded, small molecule-controlled kill switches and could inhibit the nuclear activity of the cancer cell after fusion. Perhaps this could be achieved by rapidly surrounding it with a deacidified autophagosome with dominant negative Stx17 [22] - so it cannot fuse with lysosomes. Or, bacterial phagocytosis could be employed. While at least some of these strategies may carry an increased risk of metastasis - traditional needle biopsies may also increase the risk of causing metastasis.

3. Conclusion

With regard to the brain and spinal cord, perhaps magnetism is all that’s required to cross the blood-brain and blood-spinal cord barrier - but perhaps not. Magnetic resonance-guided motorized transcranial ultrasound [23] could be used to open the blood-brain barrier at the tumor locale(s). The same technology is applicable to the spinal cord as well; MRI-guided ultrasound has already been tested in that context [24]. Instead of magnetism, perhaps a prodrug version of deschloroclozapine [25] can be employed that is blood-brain and blood-spinal cord barrier—permeable. It would be activated by an extracellular enzyme that is abundant in the brain and/or spinal cord [26] and serve as a chemoattractant for the macrophages [27]. This could be superior to magnetism because it might induce more crawling along the barrier endothelial cells until an infiltration point can be found. The question of egress from the central nervous system is also an issue. Chemotaxis and/or magnetotaxis may again be sufficient. Alternatively, maybe in the future, the CAR-Ps can somehow be directed to the lymphatic system in the brain and spinal cord and drain into a more easily accessible lymph node [28] for needle-based extraction.
This magnetism-based approach is also suitable for the macrophage-mediated delivery of oncolytic vectors to tumors that would be difficult to reach via needle, of course. This has already been accomplished in mice [4]. With regard to OVERCOME, vector replication in the carrier macrophages would be induced via small molecule, heat, or even magneto-mechanical actuation [29] once they have reached the tumor(s). Excess iron oxide nanoparticle deposition in the body from carrier macrophage lysis may not be ideal, but they should be degraded eventually [30].

Acknowledgments

The figure in this piece was created with BioRender.com.

References

  1. Renteln, M. Conditional replication of oncolytic viruses based on detection of oncogenic mRNA. Gene Therapy 2018;25(1):1–3; [CrossRef]
  2. Renteln, M. Correction: Conditional replication of oncolytic viruses based on detection of oncogenic mRNA. Gene Ther 2021;28(7):469–469; [CrossRef]
  3. Renteln, MA. Promoting oncolytic vector replication with switches that detect ubiquitous mutations. CCTR 2023;19; [CrossRef]
  4. Muthana M, Kennerley AJ, Hughes R, et al. Directing cell therapy to anatomic target sites in vivo with magnetic resonance targeting. Nature Communications 2015;6(1):8009; [CrossRef]
  5. Dowdell A, Paschke PI, Thomason PA, et al. Competition between chemoattractants causes unexpected complexity and can explain negative chemotaxis. Current Biology 2023;33(9):1704-1715.e3; [CrossRef]
  6. Zhang X, Ren L, Wu J, Feng R, Chen Y, Li R, et al. ARHGEF37 overexpression promotes extravasation and metastasis of hepatocellular carcinoma via directly activating Cdc42. J Exp Clin Cancer Res 2022;41:230. [CrossRef]
  7. Morrissey MA, Williamson AP, Steinbach AM, et al. Chimeric antigen receptors that trigger phagocytosis. Cooper JA. ed. eLife 2018;7:e36688; [CrossRef]
  8. Ito A, Teranishi R, Kamei K, et al. Magnetically triggered transgene expression in mammalian cells by localized cellular heating of magnetic nanoparticles. Journal of Bioscience and Bioengineering 2019;128(3):355–364; [CrossRef]
  9. Bausch-Fluck D, Hofmann A, Bock T, et al. A Mass Spectrometric-Derived Cell Surface Protein Atlas. PLOS ONE 2015;10(4):e0121314; [CrossRef]
  10. Bausch-Fluck D, Goldmann U, Müller S, et al. The in silico human surfaceome. PNAS 2018;115(46):E10988–E10997; [CrossRef]
  11. Zhu I, Liu R, Garcia JM, et al. Modular Design of Synthetic Receptors for Programmed Gene Regulation in Cell Therapies. Cell 2022;185(8):1431-1443.e16; [CrossRef]
  12. Kinchen JM, Ravichandran KS. Phagosome maturation: going through the acid test. Nat Rev Mol Cell Biol 2008;9(10):781–795; [CrossRef]
  13. 3 Wang X, Maxwell KG, Wang K, Bowers DT, Flanders JA, Liu W, et al. A nanofibrous encapsulation device for safe delivery of insulin-producing cells to treat type 1 diabetes. Sci Transl Med 2021;13:eabb4601. [CrossRef]
  14. Li H, Pohler U, Strehlow I, et al. Macrophage precursor cells produce perforin and perform Yac-1 lytic activity in response to stimulation with interleukin-2. Journal of Leukocyte Biology 1994;56(2):117–123; [CrossRef]
  15. Tamang DL, Alves BN, Elliott V, et al. Regulation of perforin lysis: Implications for protein disulfide isomerase proteins. Cell Immunol 2009;255(1–2):82–92; [CrossRef]
  16. Heusel JW, Wesselschmidt RL, Shresta S, et al. Cytotoxic lymphocytes require granzyme B for the rapid induction of DNA fragmentation and apoptosis in allogeneic target cells. Cell 1994;76(6):977–987; [CrossRef]
  17. Voskoboinik I, Smyth MJ, Trapani JA. Perforin-mediated target-cell death and immune homeostasis. Nat Rev Immunol 2006;6(12):940–952; [CrossRef]
  18. 8 Gupta M, Shin D-M, Ramakrishna L, Goussetis DJ, Platanias LC, Xiong H, et al. IRF8 directs stress-induced autophagy in macrophages and promotes clearance of Listeria monocytogenes. Nat Commun 2015;6:6379. [CrossRef]
  19. 9 Shiratori T, Suzuki S, Kakizawa Y, Ishida K. Phagocytosis-like cell engulfment by a planctomycete bacterium. Nat Commun 2019;10:5529. [CrossRef]
  20. 0 Kolinko I, Lohße A, Borg S, Raschdorf O, Jogler C, Tu Q, et al. Biosynthesis of magnetic nanostructures in a foreign organism by transfer of bacterial magnetosome gene clusters. Nature Nanotech 2014;9:193–7. [CrossRef]
  21. Xu C, Ren X-H, Han D, et al. Precise Detection on Cell–Cell Fusion by a Facile Molecular Beacon-Based Method. Anal Chem 2022;94(49):17334–17340; [CrossRef]
  22. Uematsu M, Nishimura T, Sakamaki Y, et al. Accumulation of undegraded autophagosomes by expression of dominant-negative STX17 (syntaxin 17) mutants. Autophagy 2017;13(8):1452–1464; [CrossRef]
  23. 3 Magnin R, Rabusseau F, Salabartan F, Mériaux S, Aubry J-F, Le Bihan D, et al. Magnetic resonance-guided motorized transcranial ultrasound system for blood-brain barrier permeabilization along arbitrary trajectories in rodents. J Ther Ultrasound 2015;3:22. [CrossRef]
  24. 4 Weber-Adrian D, Thévenot E, O’Reilly MA, Oakden W, Akens MK, Ellens N, et al. Gene delivery to the spinal cord using MRI-guided focused ultrasound. Gene Ther 2015;22:568–77. [CrossRef]
  25. 5 Nagai Y, Miyakawa N, Takuwa H, Hori Y, Oyama K, Ji B, et al. Deschloroclozapine, a potent and selective chemogenetic actuator enables rapid neuronal and behavioral modulations in mice and monkeys. Nat Neurosci 2020;23:1157–67. [CrossRef]
  26. 6 Yevtodiyenko A, Bazhin A, Khodakivskyi P, Godinat A, Budin G, Maric T, et al. Portable bioluminescent platform for in vivo monitoring of biological processes in non-transgenic animals. Nat Commun 2021;12:2680. [CrossRef]
  27. 7 Park JS, Rhau B, Hermann A, McNally KA, Zhou C, Gong D, et al. Synthetic control of mammalian-cell motility by engineering chemotaxis to an orthogonal bioinert chemical signal. Proc Natl Acad Sci U S A 2014;111:5896–901. [CrossRef]
  28. 8 Xu J-Q, Liu Q-Q, Huang S-Y, Duan C-Y, Lu H-B, Cao Y, et al. The lymphatic system: a therapeutic target for central nervous system disorders. Neural Regen Res 2022;18:1249–56. [CrossRef]
  29. 9 Beltran-Huarac J, Yamaleyeva DN, Dotti G, Hingtgen S, Sokolsky-Papkov M, Kabanov AV. Magnetic Control of Protein Expression via Magneto-mechanical Actuation of ND-PEGylated Iron Oxide Nanocubes for Cell Therapy. ACS Appl Mater Interfaces 2023;15:19877–91. [CrossRef]
  30. Mazuel F, Espinosa A, Luciani N, et al. Massive Intracellular Biodegradation of Iron Oxide Nanoparticles Evidenced Magnetically at Single-Endosome and Tissue Levels. ACS Nano 2016;10(8):7627–7638; [CrossRef]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.
Copyright: This open access article is published under a Creative Commons CC BY 4.0 license, which permit the free download, distribution, and reuse, provided that the author and preprint are cited in any reuse.
Prerpints.org logo

Preprints.org is a free preprint server supported by MDPI in Basel, Switzerland.

Subscribe

© 2024 MDPI (Basel, Switzerland) unless otherwise stated