Stimulus-triggered acquisition of pluripotency

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Stimulus-triggered acquisition of pluripotency (also known as STAP) was an alleged method of generating pluripotent stem cells by subjecting ordinary cells to certain types of stress, such as the application of a bacterial toxin, submersion in a weak acid, or physical squeezing.[1][2] The technique gained prominence in January 2014 when research by Haruko Obokata was published in Nature. Over the following months, all scientists who tried to duplicate her results failed and suspicion arose that Obokata's results were due to error or fraud. An investigation by her employer, RIKEN, was launched. On 1 April 2014, RIKEN concluded that Obokata had falsified data to obtain her results.[3] On 4 June 2014, Obokata agreed to retract the papers.[4] On August 5th, 2014, Yoshiki Sasai—Obokata's supervisor at RIKEN and one of the coauthors on the STAP cell papers—was found dead at a RIKEN facility after an apparent suicide by hanging.[5]

STAP would have been a radically simpler method of stem cell generation than previously researched methods as it would have required neither nuclear transfer nor the introduction of transcription factors.[6]

Overview[edit]

Haruko Obokata claimed that STAP cells were produced by exposing CD45+ murine spleen cells to acidic medium with a pH of 5.7 for half an hour.[6][7] Following this treatment, the cells were verified to be pluripotent by observing increasing levels of Oct-4 (a transcription factor expressed in embryonic stem cells) over the following week using an Oct4-GFP transgene.[6][8] On average only 25% of cells survived the acid treatment, but over 50% of those that survived converted to Oct4-GFP+CD45- pluripotent cells.[6] The researchers also found that treatment with bacterial toxins or physical stress were conducive to the acquisition of pluripotent markers.[1] STAP cells injected into mouse embryos grew into a variety of tissue and organs found throughout the body. According to the researchers, the chimaeric mice "[appeared] to be healthy, fertile, and normal" after one-to-two years of observation.[9] Additionally, these mice produced healthy offspring, thereby demonstrating germline transmission which is "a strict criterion for pluripotency as well as genetic and epigenetic normality."[6]

STAP cells are able to differentiate into placental cells, meaning they are more potent than embryonic stem cells or induced pluripotent stem cells (iPS).[1] It is not clear why ordinary cells do not convert into stem cells when subjected to similar stimuli under ordinary conditions, such as acidity in the body; it is hypothesized that in vivo inhibitory mechanisms may block conversion to pluripotency.[6] Research is underway to generate stimulus-triggered acquisition of pluripotency (STAP) cells using human tissue: in February 2014, Charles Vacanti and Koji Kojima (Harvard researchers originally involved in the discovery and publication of STAP) claimed to have preliminary results of STAP cells generated from human fibroblasts, but concomitantly cautioned that these preliminary results require further analysis and validation.[10]

History[edit]

In the early 2000s, Charles and Martin Vacanti conducted studies that led them to the idea that stem cells could be spontaneously created when ordinary tissues are stressed via mechanical injury or increased acidity.[11]

The technique for producing STAP cells was subsequently developed by Obokata at the Brigham and Women's Hospital (BWH), while she was studying under Charles Vacanti, and then at the RIKEN Center for Developmental Biology in Japan.[12][13] In 2008, while working at Harvard Medical School, she verified at the request of Charles Vacanti that some of the cultured cells she was working with shrank to the size of stem cells after being squeezed through a capillary tube.[1][9] She went on to test the effects of various stimuli on cells. After perfecting her technique, Obokata was able to show that white blood cells from newborn mice could be transformed into cells that behaved much like stem cells. She repeated the experiment with other cell types including brain, skin, and muscle cells with the same result.[9]

Initially Obokata's findings were met with skepticism, even among her coworkers. "Everyone said it was an artefact – there were some really hard days", she recalled.[1] The manuscript describing the work was rejected multiple times before its eventual publication in the journal Nature.[9] A series of experiments, first turning a mouse embryo green by fluorescently tagging STAP cells, then video taping the transformation of T-cells into pluripotent cells, finally convinced skeptics that the results were real.[1]

In February 2014, Vacanti said he believes the team now has evidence the same technique can be used to get stem cells from human skin cells. Early evidence suggest that the technique has also generated stem cells from human dermal fibroblasts and that there is a parallel process going on when the stem cells are generated from skin tissue.[11]

Investigation into disputed claims[edit]

In the months after the two Nature papers [9] were released, all scientists who tried to duplicate Obokata's results failed and suspicion arose that her results were due to error or fraud. An investigation into alleged irregularities was launched by RIKEN on 15 February 2014. The allegations question the use of seemingly duplicated images in the papers, and report failure to reproduce her results in other prominent stem-cell laboratories. Nature also announced they are investigating. Several stem-cell scientists defended Obokata or reserved their opinion while the investigation was ongoing.[14] To address the problem of reproducibility in other laboratories, Obokata published some technical 'tips' on the protocols on 5 March while promising the detailed procedure will be published in due course.[15]

On 11 March, Teruhiko Wakayama, one of Obokata's coauthors urged all the researchers involved to withdraw the articles, citing many "questionable points".[16] Charles Vacanti said he opposed their retraction and posted details about how to create STAP cells on his own website.[17]

On 14 March, RIKEN released an interim report of the investigation. Out of the six items being investigated, the committee concluded that there was inappropriate handling of data on two items, but did not judge the mishandling as research misconduct.[18] On 1 April, RIKEN concluded that Obokata had engaged in "research misconduct", falsifying data on two occasions. The co-authors were cleared of misconduct, but bore "grave responsibility" for not verifying the data themselves. RIKEN also announced an internal group has been established to verify whether the ‘stimulus-triggered acquisition of pluripotency’ is reproducible. [19] Obokata maintained her innocence and said she would appeal the decision.[3] On 4 June 2014, Obokata agreed to retract both papers. [4] It was officially retracted on 2 July 2014. An article analyzing the controversy, concluded that while issues of image manipulation, duplication and plagiary were potentially detectable, the reviewers could not have concluded the article was the product of academic misconduct prior to acceptance. [20]

In the wake of the controversy, observers, journalists, and former members of RIKEN have stated that the organization is riddled with unprofessional and inadequate scientific rigor and consistency, and that this is reflective of serious issues with scientific research in Japan in general.[21][22]

RIKEN commissioned a team of scientists to attempt to verify Obokata's original results and asked Obokata to participate in the effort. On 5 August 2014, Obokata's supervisor and co-author of the original paper, Yoshiki Sasai, was discovered dead by apparent suicide by hanging in a building at the RIKEN facility in Kobe, Japan.[23]

Implications[edit]

If proven to be valid, stimulus-triggered pluripotency cells could be generated more easily and efficiently compared to the existing iPS techniques.[1] If the findings can be duplicated in human tissue, it could lead to cheap and simple procedures to create patient-specific stem cells. Stem-cell researcher Dusko Ilic of King's College London called STAP cells "a major scientific discovery that will be opening a new era in stem-cell biology".[9] Shinya Yamanaka, a pioneer of iPS research, called the findings "important to understand nuclear reprogramming ... [and] a new approach to generate iPS-like cells".[1] The fact that STAP cells can form placental tissue could make the use of the cells considerably easier by bypassing the need for a donor egg and in vitro cultivation.[1]

Previously, stem cells have been created by the controversial technique of cloning embryos, or via genetic manipulation of adult cells into iPS cells. Progress on iPS-based therapies has been slow due to regulatory hurdles surrounding genetic manipulation.[9] Additionally, iPS techniques have an observed efficiency of around 1%, significantly lower than the claimed efficiency of STAP.[1]

See also[edit]

References[edit]

  1. ^ a b c d e f g h i j Cyranoski, David (2014-01-29). "Acid bath offers easy path to stem cells". Nature News (Nature Publishing Group). Retrieved 2014-01-30. 
  2. ^ Gallagher, James (2014-01-29). "Stem cell 'major discovery' claimed". BBC News. Retrieved 2014-01-31. 
  3. ^ a b Kameda, Masaaki; Otake, Tomoko (1 April 2014). "Obokata falsified data in STAP papers: probe". The Japan Times. Retrieved 2 April 2014. 
  4. ^ a b Elaine Lies (4 June 2014). "Japan researcher agrees to withdraw disputed stem cell paper". Reuters. Retrieved 4 June 2014. 
  5. ^ "STAP paper co-author Sasai commits suicide". The Japan Times. Retrieved 5 August 2014. 
  6. ^ a b c d e f Obokata, Haruko; et al. (2014-01-30). "Stimulus-triggered fate conversion of somatic cells into pluripotency". Nature 505 (7485): 641–647. doi:10.1038/nature12968. PMID 24476887. Retrieved 7 March 2014. 
  7. ^ NHS Choices (2014-01-30). "Breakthrough in stem cell creation using acid bath - What did the research involve?". nhs.co.uk. Retrieved 2014-02-06. "They put them in a weak acid solution (pH 5.7) for 30 minutes at 37°C, and then put them into petri dishes and grew them at normal pH." 
  8. ^ Thomson, Helen (2014-01-29). "Stem cell power unleashed after 30 minute dip in acid". New Scientist. Retrieved 2014-01-31. 
  9. ^ a b c d e f g Sample, Ian (2014-01-29). "Simple way to make stem cells in half an hour hailed as major discovery". The Guardian. Retrieved 2014-01-31. 
  10. ^ Thomson, Helen. "Extraordinary stem cell method tested in human tissue". New Scientist. New Scientist. Retrieved 7 March 2014. 
  11. ^ a b Connor, Steve (2014-02-09). "Exclusive: The miracle cure - scientists turn human skin into stem cells". independent.co.uk. Retrieved 2014-02-09. 
  12. ^ Grens, Kerry (29 January 2014). "New Method for Reprogramming Cells". The Scientist. 
  13. ^ "STAP cell pioneer nearly gave up on her research". The Asahi Shimbun. 30 January 2014. Archived from the original on 30 January 2014. 
  14. ^ Cyranoski, David (17 February 2014). "Acid-bath stem-cell study under investigation". Nature. doi:10.1038/nature.2014.14738. Retrieved 20 February 2014. 
  15. ^ Haruko Obokata, Yoshiki Sasai and Hitoshi Niwa (March 2014). Essential technical tips for STAP cell conversion culture from somatic cells. Nature Protocols Discussion Forum
  16. ^ "Prof. wants STAP findings withdrawn". The Yomiuri Shimbun. 11 March 2014. Retrieved 17 March 2014. 
  17. ^ Charles A Vacanti (2014)PROTOCOL FOR GENERATING STAP CELLS FROM MATURE SOMATIC CELLS. CENTER FOR TISSUE ENGINEERING & REGENERATIVE MEDICINE.
  18. ^ Press Release (14 March 2014). "Interim report on the investigation of the Obokata et al. articles". RIKEN. Retrieved 17 March 2014. 
  19. ^ Press Release (1 April 2014). "Report on STAP Cell Research Paper Investigation". RIKEN. Retrieved 2 June 2014. 
  20. ^ Cyranoski, David. "Research integrity: Cell-induced stress". Nature. Retrieved 5 August 2014. 
  21. ^ Otake, Tomoko, "‘STAPgate’ shows Japan must get back to basics in science", Japan Times, 21 April 2014
  22. ^ Schreiber, Mark, "Ongoing Obokata story seeks out scandal", Japan Times, 5 July 2014, p. 19
  23. ^ Kyodo News, "STAP paper co-author Sasai commits suicide", Japan Times, 6 August 2014, p. 1

Further reading[edit]

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