Selling Your Significance

Example From:
National Institutes of Health (NIH)

In those lucky cases where subject the matter merit is relatively obvious and easy to grasp, the key is in taking the right approach. When I say obvious, I refer to something such as curing a disease, which generally does not require a great deal of discussion to understand as a worthy end goal.

In many NIH proposals, where the end goal is saving human lives, or reducing harm, the overarching motivation is as clear as the path uncertain. In such a case, what truly makes a project significant is the credibility of its argument to advance the field in a direction. NIH proposals are therefore especially effective illustrations of a style of argument in which many people work towards a shared priority, with the divergence being in the trails they propose to blaze.

I would recommend reading the following example regardless of whether one plans on submitting to the NIH. That tight focus on picking the right approach, and understanding the manner in which they communicate that—how, for instance, do they contextualize the literature, the related progress and competing approaches; how do they mark themselves as different, while taking the best lessons of researchers at the forefront of the field? The lessons can easily be generalized to most proposal writing.

a. A literature of false starts, and a likely path (Myler Parsons R21 #1)

The following discusses the second paragraph of a section on significance. You will have to trust that, in the first, the authors did a good job of establishing significance in regards to the stated question, which regards the unknown manner in which trypanosomatid gene expression gives rise to its different stage‐specific phenotypes. Suffice to say that it matters, and the authors quickly bridged in the first paragraph from the general merit of tackling trypanosamid (the parasite responsible for what is known as African sleeping sickness) to the actionable idea that we need to know more about how its stage-specific phenotypes come about. With closing this knowledge gap as a goal and early takeaway, the questions are practical, the flow logical. What do we know? Where do we start? What proceeds is an argument by exclusion, leading to a likely path forward.

Exactly how trypanosomatid gene expression gives rise to the different stage‐specific phenotypes is not well understood. In contrast to mammals and fungi, the relative contribution of gene‐specific transcriptional control appears low in trypanosomatids, since the protein‐coding genes are organized in large polycistronic clusters [4‐8], and mRNAs derived from the same transcriptional unit vary widely in abundance within and between developmental stages [9‐11]. Differences in post‐transcriptional processing and stability play major roles in regulating mRNA abundances in T. brucei and Leishmania [12‐16], but comparison of microarray and proteomic analyses showed that there is a poor correlation between mRNA and protein abundance during Leishmania development [17]. Indeed, our own study [18] using the same biological samples from several time points during promastigote‐to‐amastigote differentiation indicated that almost a third of the 902 genes analyzed showed poor (or negative) correlation between changes in mRNA and protein levels (Fig. 1). Several studies, including our own, have provided evidence for translational control of stage‐regulation in Leishmania [19,20] and T. brucei [13,21,22]. We have also shown that stumpy form T. brucei have a paucity of polysomes [23], indicating developmental changes in translational capacity. These findings suggest that translational controls may figure prominently in modulating protein abundance in trypanosomatids. Similar layered controls have been observed in other organisms, such as Saccharomyces, in which the correlation of mRNA to protein abundance can be less than 0.2 [24].

Thus, after two paragraphs, we have approached closer and closer to the actual approach. The manner in which it does so in this paragraph is a nice sort of argument, and worth breaking down. All proposals, by definition, venture into the unknown. Yet the thing about the unknown is that one cannot say very much about it. So, here as elsewhere, the strategy is to discuss the surrounding knowns. Here, the example of mammals and fungi would suggest a certain answer to fill the gap, yet fundamental characteristics of the Trypanosomatidae family seems to rule out that path. This is one likely outcome that a reviewer might have suggested, ruled out. Another intuitive answer, "differences in post-transcriptional processing," is suggested to be equally unlikely, based both on the general literature and the work of the proposal authors. Here is another likely outcome, ruled out. Another likely answer is translational control. And here the authors find nothing to contradict. All the evidence suggests that this is promising stuff.

The second paragraph, having begun with a knowledge gap, has quickly whittled it down to a more manageable shape. The speed that it travels suggests that the authors do not feel these points are particularly controversial, but rather a matter of getting a basic premise out there in order to be certain that everyone is working from the same general assumptions. They also mimic the thought process of someone coming into their field: a few credible explanations, several ruled out, and then a decent pile of evidence implying that further efforts should be directed here. Now, let's imagine that the reviewers accept the work done in this second paragraph: this is where we should be looking, the right area of inquiry. If we accept that, what is the natural question? Where does the logical flow take us?

b. How to proceed, and why we haven't been able to proceed until now (Myler Parsons R21 #2)

The question of how to proceed is one that is deeply rooted in the state of things right now. The following is worth reading in its own right, but particularly as an example of the manner in which Why NOW? is such a fundamental question to answer in a proposal. If something is too innovative, it becomes unfeasible. If it is too conventional, someone else has probably done it, or perhaps it just wasn't worth doing. In the following paragraph, look for key words and phrases like "have begun to," "recent developments," "next generation," "exciting technological advance," "one step closer," and even the opening word of "while," which is itself a word related to time and simultaneity. This constant attention to how recently all of these things have come about seems designed to preempt a number of the natural skeptical questions mentioned in the introduction to this module. This project hasn't been done because it could only have been done right now.

While recent proteomic analyses have begun to elucidate some developmentally regulated proteins in trypanosomatids [17,25‐27], this approach remains challenging for low abundance proteins, precluding global analysis. However, recent developments allow the global measurement of the translational status of mRNAs by profiling, at the codon level, their occupation with ribosomes [24]. Ribosomes assembled onto mRNA protect about 28 nt from exogenous nuclease. Thus, if polysomal RNA is treated with nuclease, the segments of mRNAs actively undergoing translation are preserved, while the rest of the mRNA is degraded. By sequencing the protected fragments, one can obtain a measure of the relative ribosome loading of the transcript, and also the location of the ribosomes along the mRNA. The power of next generation sequencing (NGS) allows this technology to penetrate to low abundance mRNAs, opening up possibilities that are not available through other approaches, such as microarray analysis of polysomal fractions. In addition, since ribosome profiling has resolution at the codon level, it enables much more precise interrogation of exactly which open reading frames (ORFs) are being actively translated at any given, rather than just revealing which mRNAs are associated with ribosomes as in the case of microarray‐based analyses. Thus, this exciting technological advance will allow us to assess for the first time the relative translation of T. brucei mRNAs as compared to their relative abundance; taking us one step closer to the major mediator of function, the protein itself. (emphasis mine)

You also can't get away from noting that the paragraph, though it draws on the literature, largely exists to describe a concept—only one citation supports the claim about "recent developments," suggesting that they are very recent. After this, the paragraph stops citing the literature and starts providing concepts, a mixture of descriptions of the basic premise of why the approach works and descriptions of how that approach will benefit us. The bolded sentence about NGS, for instance, has a nice amplifying appeal, clearly delineating the fundamental benefit of leaning on NGS in a manner that competing approaches do not. As an R21 project, which is more experimental, the focus on innovation is appropriate and expected: the story told lines up with the grant opportunity.

In the closing sentence, we can see a broader view of the project goals. The end of this particular project does not mean the end of progress. Rather, it fits into a chain of forward movement. It is a narrative born out of the literature, sustained by present innovation, and delivered towards a shared end goal. Their methods are innovative, but they fit into a larger story. Being able to contextualize one's project as only one piece of a puzzle lends an appropriate intellectual humility (in direct repudiation to breathless salesmanship), but also promises something greater in the end. After all, if there is a whole field of people working towards this goal, there is a whole field of people who will grab onto any forward progress and in this way amplify it. This is a better sort of salesmanship, in that it strikes the reader as true.

c. An explicit acknowledgment of the inverse relationship between success and innovation (Karplus)

In the previous excerpt, we got just a hint of an important point for those proposals that seek to innovate—that any discussion of innovation is also one of feasibility. This quote, drawn from another proposal, is an explicit acknowledgment of that same point. It does not need a great deal of discussion, so here goes!

This applicant has...decades of experience with the computational methods to be used in the study, and continues to oversee the development of novel simulation methodologies. Thus, the proposed study, in spite of its novelty, has an excellent chance of success in the two-year time period. (emphasis mine)

Given that NIH proposals have a dedicated section on innovation, it is not a coincidence that this comment appeared here. While this module will generally focus on significance, background and motivation, along with the subjects that it is coupled to and the stories that unite them, it is important to get in the habit of thinking in this manner of preempting skeptical questions regardless of the section. There is no place that one shouldn't be thinking of the reader!

d. A Biased Literature (Starnbach)

A common sort of argument is one of inattention and misplaced perspectives. It tends to be dominated by words such as "myopia" and "bias," and will probably be evaluated by reviewers whose work might fall into that myopia, making it a hard needle to guide. Yet, in milder forms, such an argument of the author providing a fresh perspective is extremely common. Here is one such example. Also, as with other such examples of the type, it tends to present a picture of a field that, though its efforts are disproportionately arrayed to favor some area over another, is not irrational in doing so. In this case, it is a bias of technological availability in one area over another.

Although most investigators studying the effects of bacteria on host cells appreciate that gene regulation and protein turnover are both key to understanding how bacteria alter host cell functions, there has a been a bias towards the study of gene regulation rather than protein stability. This bias is a result of the availability of arrays and other tools that allow the simultaneous measurement of thousands of individual host cell gene transcripts during a particular stage of cell infection. Although proteomics is an active area of investigation, the ability to monitor changes in protein abundance globally is limited. Approaches to study protein levels usually involve pulse-chase experiments, the administration of protease inhibitors followed by biochemical analysis, or biochemically trapping individual protein-protein interactions. These approaches are inherently limited in the number of proteins that can be monitored at one time, and are not scalable to make them more "global."

Naturally, if the root cause of the bias is in technology, one can imagine where the rest of the proposal is going: technological innovation. But one could imagine the same argument in a humanities proposal being driven not by methodology but, say, archival availability, or unique groundwork of some sort. We will see more than one example of this in the NEH section linked to later on, including some very strong claims of bias.

e. An Incoming Threat (Dow #1)

I mentioned at the open a detail that, for me, defines much of what makes NIH proposals distinct—in the general universality of many of their ultimate goals, they can often compress their discussion of that underlying motivation in favor of bridgebuilding, from a general goal shared by many researchers to more particular research objectives and aims. If it is a fatal disease that afflicts a large number of people, the questions very quickly become not about whether the subject matter is worthwhile, but whether you can act to move things, however incrementally, towards achieving some shared goal. Yet this compression of merit is not a universal rule.

This excerpt paints a picture of a threat that mixes the present and the hypothetical. It does so through a sense of movement, a threat bearing in on the United States. Thus the merit of the general subject matter is somewhat longer than the often rote opening sentences of many NIH proposals (i.e. listing the number of people who die of the disease being studied). In order to track the movement of this passage from start to finish, I'm going to post it whole, then follow with sentence by sentence commentary. For a snapshot view, skip all that and just compare the opening sentence of this paragraph to its close—the movement from "a saprophyte in soil and water" to "a dangerous bacterial pathogen with high potential for spread into...the U.S." It is this difference between open and close that lends urgency to the project. Everything in between those two sentences exist to propel the narrative from that first, neutral phrasing to the latter, charged version.

Along the way, two main categories of sentence exist—first, those that expand and move the scope of the threat closer to home, and second, those that characterize Burkholderia pseudomallei as a threat in the first place, whether by describing fatality rates or its intractability before many antimicrobials.

Burkholderia pseudomallei (Bp) infection is a Gram-negative bacterial pathogen that normally survives as a saprophyte in soil and water, but is also capable of infecting most mammals and causing serious infections (105). Bp infection is a major cause of bacterial sepsis and chronic disseminated infections (meliodosis) in humans in Thailand and northern Australia (4-8). The fatality rate for patients with Bp infection, even with prompt and aggressive treatment, still ranges from 20% to over 50%... Moreover, Bp is an emerging pathogen and infections have been increasingly reported in many regions of the world, including Central and South America (9-13). In fact, Bp infection is now considered endemic in regions of China and India, and in Brazil (11, 12, 14). Infections with Bp are particularly dangerous because the organism is intrinsically resistant to many antimicrobials, can persist for years in the soil and in water, and can cause a wide array of clinical symptoms, ranging from acute sepsis, to chronic recurrent infection, to clinically silent infection (5-8, 15-17). Meliodosis is also an increasing problem in travelers who have visited regions of the world where Bp is endemic (18). Thus, Bp is a dangerous bacterial pathogen with high potential for spread into new regions of the world including the U.S. via deliberate or accidental introduction in soil, food, or water.

Burkholderia pseudomallei (Bp) infection is a Gram-negative bacterial pathogen that normally survives as a saprophyte in soil and water, but is also capable of infecting most mammals and causing serious infections (105).

The first sentence contains movement in itself, from soil and water to mammals with serious infections. This sets the tone for the paragraph, in moving in the direction of threats and vulnerabilities. The question is raised: what sort of infections, and what sort of mammals? The answer comes immediately.

Bp infection is a major cause of bacterial sepsis and chronic disseminated infections (meliodosis) in humans in Thailand and northern Australia (4-8).

The soil and water of the first sentence has now given way entirely to specific conditions in humans.

The fatality rate for patients with Bp infection, even with prompt and aggressive treatment, still ranges from 20% to over 50%...

Bad. This is a pathogen of severe consequences and high signifiance.

Moreover, Bp is an emerging pathogen and infections have been increasingly reported in many regions of the world, including Central and South America (9-13).

The scope continues to expand: suddenly it is popping up on different continents entirely. It's not pointed out explicitly, but if it can hop from Asia/Australia to South and Central America, what is to stop its spread to North America?

In fact, Bp infection is now considered endemic in regions of China and India, and in Brazil (11, 12, 14).

A step worse still. It is spreading and entrenching.

Infections with Bp are particularly dangerous because the organism is intrinsically resistant to many antimicrobials, can persist for years in the soil and in water, and can cause a wide array of clinical symptoms, ranging from acute sepsis, to chronic recurrent infection, to clinically silent infection (5-8, 15-17).

Like the previous sentence on fatality rates, this sentence serves to collect all the reasons that Bp is particularly dangerous. It is not just spreading and entrenching but difficult to diagnose, resistant to treatment, and long-lasting in various reservoirs of soil and water.

Meliodosis is also an increasing problem in travelers who have visited regions of the world where Bp is endemic (18).

We return to the idea of an infection spreading on networks of travel.

Thus, Bp is a dangerous bacterial pathogen with high potential for spread into new regions of the world including the U.S. via deliberate or accidental introduction in soil, food, or water.

The pathogen arrives in the US.

The paragraph itself bears in on the U.S. Altogether, this calls on several of the argument styles mentioned in the introduction. But the fundamental point is best grasped by seeing what comes next.

f. An Underrated Threat (Dow #2)

The previous paragraph established the threat to the United States to be greater than first glance might suggest. This paragraph further amplifies that threat, suggesting that the literature has missed a key point. In the process, it tells two stories. The first story is about the nature of Bp. The second story is about how the field understands Bp. In between those two is a gap, and this work takes place in that gap.

Most proposals are similar: there is always a distance between our intellectual artifices and the underlying reality, and research projects, at least the well-designed ones, tend to bring us closer to that underlying reality. Reviewers know that, but do they trust you to be the one to advance the field? There is absolutely no way to make that argument without knowing what the field is up to.

Let's take a look at the manner in which they do so, the mixture of personal communications and patterns of fact.

Currently Bp is not considered a primary enteric pathogen for infection of humans. At present, infection with Bp is presumed to occur following inhalation or cutaneous inoculation, though the actual link between cutaneous exposure and infection is weak (Dr. Sharon Peacock, see Letter of Support). Thus, current treatment and prevention efforts for human meliodosis do not consider the impact of oral infection or persistent fecal carriage and shedding of the organism (5, 7, 8). There is however epidemiological evidence to suggest that oral infection with Bp does occur in humans. For example, outbreaks of meliodosis in villages in Indonesia have been linked directly to drinking water supplies contaminated with Bp, which can survive for years in water (10). Infections with Bp increase significantly during times of greater exposure to very wet conditions (eg, rice farming during the monsoon season), which would be consistent with oral exposure to a water borne agent (20-22). Outbreaks of meliodosis have also been associated with tsunami events (23, 24). In addition, patients with meliodosis have been misdiagnosed as having typhoid (enteric fever)(25). In fact, clinical observations (Dr. Peacock, personal communication) suggest that oral infection may be a much more important route of infection with Bp than previously assumed. Since Bp can persist in water or soil for years, enteric infection of humans with Bp would have major public health consequences (26, 27).

The extent of private correspondence is interesting. It is also worth noting that there is content beyond this—the paragraphs that succeed it describe the author's own experience demonstrating a highly enteric nature of the pathogen in their work with mice. They first noted this quality without even looking for it; it was such a strong tendency that it just kept happening to them. They then followed it up with a more systematic examination, demonstrating that the enteric infection was "not only a property of laboratory adapted strains of Bp," but also of Bp isolates from medical clinics. Ultimately they developed a "mouse model of enteric Bp infection" that is entirely novel and would serve as the foundation for the present project.

In short, there is significant groundwork described, all of it presented within the opening section on significance. And all serves to begin separating the understanding of the authors from what is said to be a misconception of the literature. There is even a reference to the fact that "current treatment and prevention efforts" do not treat it as enteric, pointing to an extremely actionable outcome of their study. Especially given the sense of a threat bearing in communicated in the first paragraph, one gets a feeling that every moment that this study is not funded perpetuates a potentially flawed and dangerous situation. Assuming that the underlying argument is convincing, the urgency is undeniable.

Contact:
Nural Akchurin, Associate Dean for Research
College of Arts & Sciences
806.834.8838
nural.akchurin@ttu.edu